Repetitive Suicidal Behaviors in a Case With a New Mutation of Wolfram Syndrome; A Jump From the Gene to The Behavior

Wolfram syndrome (WS) is a rare autosomal recessive neurodegenerative disease with variable symptoms including neuropsychiatric manifestations. A 26-year-old man was reported with classic symptoms of WS and repetitive psychiatric hospitalizations and at least 16 suicidal attempts. The genetic study demonstrated a novel homozygous stop-codon mutation on the WFS1 gene. This special type of mutation may be related to repetitive suicidal behaviors in this case of WS. Psychological support should be a routine practice in patients with WS.

Comparison of Interpretation between Pyrosequencing and Xpert MTB/RIF Assay in Multidrug-Resistant Tuberculosis

Indonesia is one of the countries with the highest multidrug-resistant tuberculosis cases in the world. Rapid molecular test using the Xpert MTB/RIF assay is one of the detection methods for MDR-TB. Early detection of MDR-TB is crucial for early initiation of treatment. However, Xpert MTB/RIF assay only detects the rpoB gene mutations associated with Rifampicin resistance. Recently, WHO recommends the use of Pyrosequencing, a DNA sequencing method that can detect not only the rpoB gene but also katG and/or inhA gene mutations associated with Isoniazid resistance. The aims of this study were to compare the interpretation between the two methods and to determine the differences in codon mutation position detection of the rpoB gene and mutation detection of the katG and/or inhA gene. This was a cross-sectional comparative observational study on patients ≥18 years old interpreted as RR-TB patients based on Xpert MTB/RIF assay results who had not received MDR-TB drugs at Dr. Hasan Sadikin General Hospital, Bandung, Indonesia. Results showed there were 40 Rifampicin-resistant TB subjects interpreted by Xpert MTB/RIF assay while Pyrosequencing interpreted 30 MDR-TB, 9 RR-TB and one Isoniazid-resistant TB subjects in January - February 2020. The detection of rpoB gene codon mutation position between Xpert MTB/RIF assay and Pyrosequencing methods was not significantly different (p=0.389). Pyrosequencing had detected 27 katG gene mutations, 3 inhA gene mutations, one katG and inhA gene mutation. To conclude, Pyrosequencing can be used for accurate detection of Rifampicin and Isoniazid resistance in MDR-TB.

An evolutionary analysis of the SARS-CoV-2 genomes from the countries in the same meridian (Preprint)

UNSTRUCTURED In the current study we analyzed the genomes of SARS-CoV-2 strains isolated from Italy, Sweden, Congo (countries in the same meridian) and Brazil, as outgroup country. Evolutionary analysis revealed codon 9628 under episodic selective pressure for all four countries, suggesting it as a key site for the virus evolution. Belonging to the P0DTD3 (Y14_SARS2) uncharacterized protein 14, further investigation has been conducted showing the codon mutation as responsible for the helical modification in the secondary structure. According to the predictions done, the codon is placed into the more ordered region of the gene (41-59) and close the area acting as transmembrane (54-67), suggesting its involvement into the attachment phase of the virus. The predicted structures of P0DTD3 mutated and not confirmed the importance of the codon to define the protein structure and the ontological analysis of the protein emphasized that the mutation enhances the binding probability.

An evolutionary analysis of the SARS-CoV-2 genomes from the countries in the same meridian

In the current study we analyzed the genomes of SARS-CoV-2 strains isolated from Italy, Sweden, Congo (countries in the same meridian) and Brazil, as outgroup country. Evolutionary analysis revealed codon 9628 under episodic selective pressure for all four countries, suggesting it as a key site for the virus evolution. Belonging to the P0DTD3 (Y14_SARS2) uncharacterized protein 14, further investigation has been conducted showing the codon mutation as responsible for the helical modification in the secondary structure. According to the predictions done, the codon is placed into the more ordered region of the gene (41-59) and close the area acting as transmembrane (54-67), suggesting its involvement into the attachment phase of the virus. The predicted structures of P0DTD3 mutated and not confirmed the importance of the codon to define the protein structure and the ontological analysis of the protein emphasized that the mutation enhances the binding probability.

A Unique Histone H3K4me3 Genome Binding Pattern Identified in a Cancer Pedigree with anMLL3Germline Mutation

Background and Objective: Evidence suggests that alterations in histone modifications are crucial in cancer development and progression. Among these, histone 3 lysine 4 trimethylation (H3K4me3) which is one of the most common histone modifications, is found in promoters of active genes and is proposed to promote gene expression. Aberrant H3K4 methylation is association with cancer. In our previous study, we found a germline MLL3 stop codon mutation in a Chinese pedigree. MLL3 proteins act as methyltransferase and regulate H3K4 methylation. Our study aims to evaluate the H3K4me3 genome binding pattern in this cancer pedigree with MLL3 mutation and have further understanding about pathogenesis in this pedigree. Also we hope to find valuable epigenetic marker of cancer. Materials and Methods: To evaluate 3 cancer patients in a Chinese pedigree with a heterozygous stop codon mutation in MLL3 (coding a histone H3K4me3 methyltransferase) segregating with colorectal carcinoma (CRC) and acute myeloid leukemia (AML) and a mutation-free normal control, ChIP-chip analyses were performed to determine which promoters coprecipitated with the H3K4me3 antibody in both patients and control. The pulled-down genes were mapped to Kyoto Encyclopedia Of Genes And Genomes (KEGG) pathways. Results:We selected 1 AML patient, 2 CRC patients, and 1 normal control from a Chinese pedigree. A previous study showed that the 3 patients all harbored an MLL3 stop codon mutation. MLL3 belongs to the human TRX/MLL family and is an important mammalian H3K4 methyltransferase. In this study, using ChIP-chip, we detected the global promoter occupancy profile of H3K4me3 in the two different tumor types compared with a normal control. Compared with the normal control, H3K4me3 differentially bound to the promoter regions of 2425 (CRC), 22319 (CRC), and 2385 (AML) genes in the three patients, with an overlap of 877 genes between AML and CRC tumors. To our surprise, the total numbers of promoters pulled down by the H3K4me3 antibody were not significantly different between the 3 cancer patients and the mutation-free normal control. However, the H3K4me3 binding patterns were unique in the cancer patients compared with the normal control. We also noticed a subtle difference in H3K4me3 binding patterns between the AML patient and the 2 CRC patients. The mother-son pair (CRC-AML) and the sister pair (CRCs) each should have 50% genetic similarities (sex chromosome genes were excluded); however, given the same MLL3 nonsense mutation, the CRC mother-AML son pair showed greater differences in H3K4me3 pulled-down genes than did the CRC sister pair. To understand the possible biological effects of H3K4me3 in AML and CRC, we annotated the functions of the identified genes by mapping them to KEGG pathways. Although the number of identified genes was not significantly differ between patients and control, we found an unique profile of H3K4me3 in AML and CRC patients compared with control that the identified genes were concentrated in the carcinogenesis-associated pathways, including the TGF-beta, Wnt, and MAPK signaling pathways. Conclusions:The results indicate that the H3K4me3 patterns are differentially altered in different cancers, which may play a role in carcinogenesis, and the MLL3 mutation may influence the alteration of H3K4 methylation. Our studies make it is possible to link genetic changes and epigenetic modifications. Clearly, a large sample cohort study is needed to test H3K4me3 binding patterns in MLL3 mutation patients, as well as in all cancer patients and even normal individuals. Disclosures No relevant conflicts of interest to declare.

PCR-RFLP for detection of Fusarium graminearum genotypes with resistance to phenamacril

Phenamacril is a cyanoacrylate fungicide that provides excellent control of Fusarium head blight (FHB) or wheat scab, which is caused predominantly by Fusarium graminearum and Fusarium asiaticum. Previous studies revealed that codon mutations of the myosin-5 gene of Fusarium spp. conferred resistance to phenamacril in vitro lab experiments. In this study, PCR-RFLP (polymerase chain reaction-restriction fragment length polymorphism) was developed to detect three common mutations (A135T, GCC to ACC at codon 135; S217L, TCA to TTA at codon 217, and E420K, GAA to AAA at codon 420) in F. graminearum induced by fungicide domestication in vitro. PCR products of 841 bp (for mutation of A135T), 802 bp (for mutation of S217L) or 1649 bp (for mutation of E420K) in myosin-5 gene were amplified respectively by appropriate primer pairs. Restriction enzyme KpnⅠ, TasⅠ or DraⅠ was used to distinguish phenamacril-sensitive and -resistant strains with mutation genotypes of A135T, S217L and E420K, respectively. KpnⅠ digested the 841 bp PCR products of phenamacri-resistant strains with codon mutation A135T into two fragments of 256 bp and 585 bp. In contrast, KpnⅠ did not digest the PCR products of sensitive strains. TasⅠ digested the 802 bp PCR products of phenamacril-strains with codon mutation S217L into three fragments of 461 bp, 287bp and 54 bp. In contrast, TasⅠ digestion of the 802 bp PCR products of phenamacril-sensitive strains resulted in only two fragments of 515bp and 287bp. DraⅠ digested the 1649 bp PCR products of phenamacril-resistant strains with codon mutation E420K into two fragments of 932 bp and 717 bp, while the PCR products of phenamacril-sensitive strains was not digested. The three genotypes of resistance mutations were determined by analyzing electrophoresis patterns of the digestion fragments of PCR products. The PCR-RFLP method was evaluated on 48 phenamacril-resistant strains induced by fungicide domestication in vitro and compared with the conventional method (mycelial growth on fungicide-amended agar). The accuracy of the PCR-RFLP method for detecting the three resistant mutation genotypes of F. graminearum to phenamacril was 95.12% compared with conventional method. Bioinformatics analysis revealed that the PCR-RFLP method could also be used to detect the codon mutations of A135T and E420K in F. asiaticum.