Genomic Profile of SARS-COV-2 Associated with COVID-19 Outbreaks in N’Djamena, Chad

Background: SARS-CoV-2 has circulated worldwide with dramatic consequences. In Chad, we have no data reported of variants. The aim of this study was to identify the SARS-CoV-2 variants that circulated during the epidemic from 2020 to 2021. Methods: This is a cross-sectional, descriptive study carried out between 2020 and 2021. Samples from patients with suspected COVID-19 were tested in five laboratories in N’Djamena. One hundred quality samples of the positives were sequenced in Kinshasa using Oxford nanopore technologies minion and the Protocol Midnight SARS-CoV2. Data were processed using Excel version 16 software. Results: Of the 100 samples sequenced, 77 (77%) produced sequences, 23 (23%) did not. The genomic profiles were wild-type Wuhan and minor mutations (19A, 19B (A), 20A (B.1, B.2), 20B (AV.1), 20D (B.1.1.1 /C.36), 20C), variant of concern Alpha (20I), variant of concern Delta (21A/J), variant of interest Eta (21D), variant of concern Omicron (21K) and unclassified variant under surveillance (B.1.640). Of these variants, the maximums were detected in patients aged 26 - 35 with 30.26% and 25.26% in 36 - 45. However, 24.67% were in travelers and 75.32% in residents, 35.06% in those vaccinated against COVID-19 and 62.33% in non-vaccinates. The estimated case-fatality rate was 2.44% (107/4374). Conclusion: This work has provided preliminary data on COVID-19 and SARS-CoV-2 variants circulating during the 2020-2021 epidemics in Chad.

Comparative genomic study of gastric epithelial cells co-cultured with Helicobacter pylori

AIM:To identify genes potentially involved in Helicobacter pylori(H.pylori)-induced gastric carcinogenesis.METHODS:GES-1 cells were co-cultured with H.pylori strains isolated from patients with gastric carcinoma(GC,n = 10) or chronic gastritis(CG,n = 10) for in vitro proliferation and apoptosis assays to identify the most and least virulent strains.These two strains were cagA-genotyped and used for further in vivo carcinogenic virulence assays by infecting Mongolian gerbils for 52 wk,respectively;a broth free of H.pylori was lavaged as control.Genomic profiles of GES-1 cells cocultured with the most and least virulent strains were determined by microarray analysis.The most differentially expressed genes were further verified using quantitative real-time polymerase chain reaction in GES-1cells infected with the most and least virulent strains,and by immunohistochemistry in H.pylori positive CG,precancerous diseases,and GC biopsy specimens in an independent experiment.RESULTS:GC-derived H.pylori strains induced a potent proliferative effect in GES-1 cells in co-culture,whereas CG-derived strains did not.The most(from a GC patient) and least(from a CG patient) virulent strains were cagA-positive and negative,respectively.At week 52,CG,atrophy,metaplasia,dysplasia,and GC were observed in 90.0%,80.0%,80.0%,90%,and 60.0%,respectively,of the animals lavaged with the most virulent strain.However,only mild CG was observed in 90% of the animals lavaged with the least virulent strain.On microarray analysis,800 differentially expressed genes(49 up-and 751 down-regulated),involving those associated with cell cycle regulation,cell apoptosis,cytoskeleton,immune response,and substance and energy metabolisms,were identified in cells co-cultured with the most virulent strain as compared with those co-cultured with the least virulent strain.The six most differentially expressed genes(with a betweenness centrality of 0.1-0.2) were identified among the significant differential gene profile network,including JUN,KRAS,BRCA1,SMAD2,TRAF1,and HDAC6.Quantitative real-time polymerase chain reaction analyses verified that HDAC6 and TRFA1 mRNA expressions were significantly more up-regulated in GES-1 cells cocultured with the most virulent strain than in those cocultured with the least virulent strain.Immunohistochemistry of gastric mucosal specimens from H.pyloripositive patients with CG,intestinal metaplasia(IM),dysplasia,and GC showed that moderately positive and strongly positive HDAC6 expression was detected in 21.7% of CG patients,30.0% of IM patients,54.5% of dysplasia patients,and 77.8% of GC patients(P < 0.001).The up-regulation of TRAF1 expressions was detected in 34.8%,53.3%,72.7%,and 88.9% specimens of CG,IM,dysplasia,and GC,respectively(P < 0.001).CONCLUSION:The overexpression of HDAC6 and TRAF1 in GES-1 cells co-cultured with the GC-derived strain and in H.pylori-positive dysplasia and GC suggests that HDAC6 and TRAF1 may be involved in H.pyloriinduced gastric carcinogenesis.

Identification of IL11RA and MELK amplification in gastric cancer by comprehensive genomic profiling of gastric cancer cell lines

AIM To identify common copy number alterations on gastric cancer cell lines.METHODS Four gastric cancer cell lines(ACP02, ACP03, AGP01 and PG100) underwent chromosomal comparative genome hybridization and array comparative genome hybridization. We also confirmed the results by fluorescence in situ hybridization analysis using the bacterial artificial chromosome clone and quantitative real time PCR analysis.RESULTS The amplification of 9p13.3 was detected in all cell lines by both methodologies. An increase in the copy number of 9p13.3 was also confirmed by fluorescence in situ hybridization analysis. Moreover, the interleukin 11 receptor alpha(IL11RA) and maternal embryonic leucine zipper kinase(MELK) genes, which are present in the 9p13.3 amplicon, revealed gains of the MELK gene in all the cell lines studied. Additionally, a gain in the copy number of IL11 RA and MELK was observed in 19.1%(13/68) and 55.9%(38/68) of primary gastric adenocarcinoma samples, respectively. CONCLUSION The characterization of a small gain region at 9p13.3 in gastric cancer cell lines and primary gastric adenocarcinoma samples has revealed MELK as a candidate target gene that is possibly related to the development of gastric cancer.

Comprehensive review on endoscopic ultrasound-guided tissue acquisition techniques for solid pancreatic tumor

Pancreatic ductal adenocarcinoma is speculated to become the second leading cause of cancer-related mortality by 2030,a high mortality rate considering the number of cases.Surgery and chemotherapy are the main treatment options,but they are burdensome for patients.A clear histological diagnosis is needed to determine a treatment plan,and endoscopic ultrasound(EUS)-guided tissue acquisition(TA)is a suitable technique that does not worsen the cancer-specific prognosis even for lesions at risk of needle tract seeding.With the development of personalized medicine and precision treatment,there has been an increasing demand to increase cell counts and collect specimens while preserving tissue structure,leading to the development of the fine-needle biopsy(FNB)needle.EUS-FNB is rapidly replacing EUS-guided fine-needle aspiration(FNA)as the procedure of choice for EUS-TA of pancreatic cancer.However,EUS-FNA is sometimes necessary where the FNB needle cannot penetrate small hard lesions,so it is important clinicians are familiar with both.Given these recent developments,we present an up-to-date review of the role of EUS-TA in pancreatic cancer.Particularly,technical aspects,such as needle caliber,negative pressure,and puncture methods,for obtaining an adequate specimen in EUS-TA are discussed.

Filtering for SNPs with high selective constraint augments mid-parent heterosis predictions in wheat(Triticum aestivum L.)

To extend the contemporary understanding into the grain yield heterosis of wheat, the current study investigated the contribution of deleterious alleles in shaping mid-parent heterosis(MPH). These alleles occur at low frequency in the genome and are often missed by automated genotyping platforms like SNP arrays. The deleterious alleles herein were detected using a quantitative measurement of evolutionary conservation based on the phylogeny of wheat and investigations were made to:(1) assess the benefit of including deleterious alleles into MPH prediction models and(2) understand the genetic underpinnings of deleterious SNPs for grain yield MPH using contrasting crosses viz. elite × elite(Exp. 1) and elite × plant genetic resources(PGR;Exp. 2). In our study, we found a lower allele frequency of moderately deleterious alleles in elites compared to PGRs. This highlights the role of purifying selection for the development of elite wheat cultivars. It was shown that deleterious alleles are informative for MPH prediction models: modelling their additive-by-additive effects in Exp. 1 and dominance as well as associated digenic epistatic effects in Exp. 2 significantly boosts prediction accuracies of MPH. Furthermore,heterotic-quantitative trait loci's underlying MPH was investigated and their properties were contrasted in the two crosses. Conclusively, it was proposed that incomplete dominance of deleterious alleles contributes to grain yield heterosis in elite crosses(Exp. 1).

Pancreatic cancer with synchronous liver and colon metastases: A case report

BACKGROUND Metastasis of pancreatic cancer to the colon is rare and the features need to be further elucidated.Herein,we report a rare case of pancreatic cancer with simultaneous liver and colon metastases.CASE SUMMARY A 48-year-old man with intrahepatic space-occupying lesions based on a computed tomography scan was admitted to our hospital for further treatment.Abdominal magnetic resonance imaging revealed a 6.4 cm×4.2 cm mass in the tail of the pancreas and multiple low-density masses in the liver parenchyma.In addition,a mass of 2.2 cm×1.6 cm with surface congestive erosions in the sigmoid colon was detected by colonoscopy.Histopathological examination of biopsies from both the liver and colon lesions revealed a moderately to poorly differentiated adenocarcinoma.Immunohistochemical staining of the colon tumor was positive for cytokeratin(CK)7 and CK,but negative for colorectal adenocarcinoma-related markers CK 20,CDX2,and SATB2,thus indicating that the metastasis originated from the pancreas.Next-generation sequencing for genomic profiling of the liver and colon metastases both found mutations in KRAS(p.G12D)and TP53(c.376-1delG),with microsatellite stable and low tumor mutational burden without actionable or cancer-predisposing gene mutations detected.The patient was subsequently treated with 12 cycles of FOLFIRINOX which led to a sustainable response,followed by ongoing maintenance treatment with irinotecan plus fluorouracil.CONCLUSION For this rare case,careful evaluation of histopathological and immunohistochemical staining results are required.The genomic profiling of colon lesions was revealed for the first time,and FOLFIRINOX showed good treatment efficacy in this patient.

The N^6-adenine methylation in yeast genome profiled by single-molecule technology

The most common and abundant DNA modification is 5-methylcytosine（5mC）,which has been well-established as an epigenetic mark regulating gene expression in eukaryotes（Jones,2012）.Another DNA modification N^6-methyldeoxyadenosine（6mA）,previously reported as a widespread DNA methylation in prokaryotes.