Sequence Databases Research Articles

Hereditary Transthyretin Amyloidosis in Israel: Genetic Landscape and Clinical Characteristics.

Hereditary transthyretin (ATTRv) amyloidosis is a rare, adult-onset autosomal-dominant disorder caused by pathogenic variants in the transthyretin (TTR) gene. Data about relevant variants in specific populations and typical initial manifestations may facilitate early diagnosis and treatment. We here describe the genetic landscape of ATTRv amyloidosis in Israel. Genetic and clinical data of TTR variant carriers and ATTRv amyloidosis patients were collected from a national referral clinic and other subspecialty clinics in Israel. Genotype-phenotype correlations of the detected variants were detailed. In addition, two large Israeli exome sequence (ES) databases were screened for TTR variants. Seven heterozygous disease-causing variants in TTR were identified among 95 adults (52 males, 50.7%). The Ser77Tyr variant was found in 68 (71.6%) subjects of Jewish Yemenite ancestry. Val122Ile was found in 9 (9.4%) subjects and was the only variant detected in individuals of Arab ethnicity. Other variants were Thr60Ala, Val30Met, Val32Ala, Ala81Val, and Glu89Val. Thirty-five individuals were ATTRv amyloidosis patients (25 males, 71.4%), diagnosed at a mean age of 62.5 ± 6.7 years, and 23 (63.7%) were due to Ser77Tyr. Initial symptoms were mostly related to carpal tunnel syndrome, and the sensitivity of scintigraphy was low for Ser77Tyr but high for Thr60Ala and Val32Ala variants. TTR pathogenic variants were detected in 14 of approximately 36,600 subjects who underwent ES, including Val122Ile in 9 subjects of Arab ethnicity. Most ATTRv amyloidosis cases in Israel are attributable to the Ser77Tyr variant. However, other variants also contribute to disease occurrence, and testing is warranted in clinically suspected patients.

Benefits of HIV-1 transmission cluster surveillance: a French retrospective observational study of the molecular and epidemiological co-evolution of recent circulating recombinant forms 94 and 132.

Molecular surveillance is an important tool for detecting chains of transmission and controlling the HIV epidemic. This can also improve our knowledge of molecular and epidemiological factors for the optimization of prevention. Our objective was to illustrate this by studying the molecular and epidemiological evolution of the cluster including the new circulating recombinant form (CRF) 94_cpx of HIV-1, detected in 2017 and targeted by preventive actions in 2018. In June 2022, 32 HIV-1 sequence databases from French laboratories were screened to identify all individuals who had acquired CRF94_cpx or a similar strain, whatever the date of diagnosis. Phylogenetic analyses were performed with the sequences identified, and biological parameters were collected at the time of diagnosis and after the start of treatment to analyse the evolution of the cluster. Full genomes were sequenced to characterize the new strains. We analysed 98 HIV-1 isolates: 63 were CRF94, three were unclassifiable, and the other 32 formed a new cluster containing a new recombinant, CRF132_94B, derived from CRF94 and a subtype B strain. At least 95% of the individuals in both the CRF94 and CRF132 clusters were men who have sex with men (MSM), most of whom had acquired HIV less than 12 months before diagnosis. The number of CRF94 diagnoses declined drastically after 2018, but CRF132 strains spread widely between 2020 and 2022, into a different area of Ile-de-France region and within a younger population nevertheless aware of pre-exposure prophylaxis. Higher viraemia, lower CD4 cell counts and delayed treatment efficacy suggested that CRF94 was more virulent than CRF132, possibly due to the F subtype fragment of the vif gene. These findings highlight the role of the MSM transmission cluster in spreading HIV and new variants. They show also the benefits of cluster surveillance for improving the targeting of preventive interventions, detecting the emergence of new strains and enriching our knowledge on virulence mechanisms. However, these investigations require support with sufficient resources dedicated to a regional or national programme to be responsive and effective.

ATAD2 is a potential immunotherapy target for patients with small cell lung cancer harboring HLA-A∗0201.

Small cell lung cancer (SCLC) represents a highly aggressive neuroendocrine tumour with a dismal prognosis. Currently, the identification of a specific tumour antigen that can facilitate immune-based therapies for SCLC remains elusive. We employed liquid chromatography-tandem mass spectrometry (LC-MS/MS) to analyse cancer/testis antigens (CTAs) in SCLC cell lines and human tumour specimens. Immunohistochemistry of clinical specimens was performed to compare protein expression in SCLC, non-small cell lung cancer (NSCLC), and matched normal-adjacent tissues. Additionally, publicly available RNA sequencing databases were interrogated to identify gene expression patterns in different SCLC subtypes and in different disease stages. Distinct numbers and types of CTAs were identified across SCLC subtypes, with significantly higher expression levels of ATPase family AAA domain-containing protein 2 (ATAD2) observed in SCLC compared to normal adjacent tissues and NSCLC tissues. A dynamic expression pattern of ATAD2 was found throughout the clinical course of SCLC and exhibited a positive correlation with achaete-scute family bHLH transcription factor 1 (ASCL1) expression in SCLC. Immunopeptidomics analysis identified the YSDDDVPSV sequence derived from the HLA-A∗02:01 restriction epitope of ATAD2 as a highly promising tumour antigen candidate for potential immunotherapy applications. YSDDDVPSV immunopeptides were confirmed to be present in SCLC-A and SCLC-N with HLA-A∗02:01 restriction. Notably, HLA-A∗02:01T cells exhibited a robust response upon stimulation with YSDDDVPSV immunopeptide pulsed by T2 cells. Our findings highlight the potential of targeting the ATAD2 YSDDDVPSV immunopeptide for SCLC immunotherapy, thereby offering a promising avenue for the development of adoptive T cell therapies to effectively treat ASCL1-positive or NEUROD1-positive SCLC carrying HLA-A∗02:01. This study was supported by the National key R&D program of China (2022YFC2505000); National Natural Science Foundation of China (NSFC) general program (82272796) NSFC special program (82241229); CAMS Innovation Fund for Medical Sciences (CIFMS 2022-I2M-1-009); CAMS Key Laboratory of Translational Research on Lung Cancer (2018PT31035); Aiyou foundation (KY201701). National key R&D program of China (2022YFC2505004). NSFC general program (81972905). Medical Oncology Key Foundation of Cancer Hospital Chinese Academy of Medical Sciences (CICAMS-MOCP2022012).

Maintaining taxonomic accuracy in genetic databases: A duty for taxonomists—Reanalysis of the DNA sequences from Mercan et al. (2024) on the genus Potamothrix (Annelida, Clitellata) in Turkish lakes

Public DNA sequence databases such as GenBank are widely used for identification of organisms in ecological and taxonomic studies. It is important that these public databases contain as few mistakes as possible and that any errors detected in these databases are reported. Here, we reanalyzed the COI sequences of Mercan et al. (2024) and showed that they were mistakenly considered by these authors as belonging to different populations (haplotypes) within the species Potamothrix hammoniensis (Tubificinae). We found that they corresponded to four distinct Tubificinae lineages (species), Pothamothrix alatus paravanicus, Potamothrix bavaricus, Tubifex sp. and Potamothrix sp. Despite these identification errors, the data from Mercan et al. (2024) remain interesting as they provide new information on the diversity of the genus Potamothrix in Turkey. Prompt measures must be taken to correct these errors and prevent them from being detrimental to future studies.

Isolation and Identification of Hydrocarbon-Degrading Bacteria from Diesel-Contaminated Soils

Aim: To Isolate, screen and identify bacterial species with hydrocarbon biodegradative potentials from diesel-polluted soils using redox indicator (2% v/v of 2,6 – dichlorophenol indophenols- DCPIP) and Turbidity measurements. Place and Duration of Study: Microbiology laboratory in the Department of Environmental Management and Toxicology, Federal University of Petroleum Resources, Effurun, between 2022 and 2024. Methodology: Soil samples from diesel-contaminated site were collected from diesel mechanic workshop, analysed using the spread plate isolation technique on Bushnell Haas mineral salt agar selective medium, were screened for Bacterial species with hydrocarbon degradative potential using the redox indicator (2% v/v of 2,6 – dichlorophenol indophenols- DCPIP) and Turbidity measurement. Selected Bacterial isolates were used for the hydrocarbon degradation studies, were characterized based on biochemical tests and identified with the aid of the Biolog database and 16S rRNA gene sequencing techniques. Results: Eight diesel degrading Bacteria species were identified, belonging to the following genera; Acinetobacter, Klebsiella, and Pseudomonas. Among these, the four most potent degraders were identified as Acinetobacter rudis (M2712942.1), Acinetobacter bereziniae (MT111619.1), Pseudomonas koreensis (MW794239.1), and Klebsiella aerogenes (CP070520.1) and the % diesel reduction achieved in 15 days were; 70%, 48%, 47% and 45%, while the % Turbidity increase were 84%, 79%, 78% and 76% respectively. Consequently, the partial 16S rRNA gene sequences revealed the two most potent diesel degrading bacterial strains were Acinetobacter rudis and Acinetobacter bereziniae with 70% and and 48% respectively. Conclusion: These findings demonstrate the potential of indigenous bacteria for effective bioremediation of petroleum-contaminated environments. The study provides critical insights into sustainable solutions for mitigating hydrocarbon pollution, contributing to advancements in environmental microbiology and ecosystem restoration.

The InBIO Barcoding Initiative Database: contribution to the knowledge of DNA barcodes of the vascular plants of north-eastern Portugal

Metabarcoding is invaluable for understanding trophic interactions, enabling high-resolution and rapid dietary assessments. However, it requires a robust DNA barcode reference library for accurate taxa identification. This dataset has been generated in the framework of the InBIO Barcoding Initiative (IBI) and Agrivole project. The integration of these two projects was crucial, as Agrivole aimed to investigate the trophic niche of small mammals in Trás-os-Montes Region through DNA metabarcoding, which required a reliable plant DNA barcode library for this same region. Given the large number of species not yet represented in international databases, a survey of local plants was essential to fill this gap. Thus, this study created an accurate DNA reference database for the plants of the Trás-os-Montes Region of Portugal. The current DNA reference database contains 632 vascular plant samples, all morphologically identified and belonging to 435 species. This represents 14% and 38.7% of the total known plant species for Portugal and the study area, respectively. Of the 1781 barcode sequences provided in this dataset, 1099 contain new information (61.7%) at different levels: 254 (13.6%, ITS2: 41, trnL-ef: 126, trnL-gh: 87) are completely new to GenBank and/or BOLD databases at the time of publication, 438 (24.6%, ITS2: 59, trnL-ef: 173, trnL-gh: 206) are new records for a given species and 407 (22.9%, ITS2: 187, trnL-ef: 206, trnL-gh: 14) provide additional information (e.g. different bp length, intraspecific genetic variability); the remaining 682 sequences (38.3%) are equal (100% identity) to sequences already publicly available for the identified species. Overall, this dataset represents a significant contribution to the genetic knowledge of vascular plants represented in public libraries. This is one of the public releases of the IBI database, which provides genetic and distributional data for several taxa. All vouchers are deposited in the Herbarium of the Museum of Natural History and Science of the University of Porto (MHNC-UP) and their DNA barcodes are publicly available in the Barcode of Life Data System (BOLD), NCBI GenBank online databases and International Nucleotide Sequence Database Collaboration (INSDC).

AdipoR1 enhances the radiation resistance via ESR1/CCNB1IP1/cyclin B1 pathway in hepatocellular carcinoma cells

Hepatocellular carcinoma is one of the most common malignant tumors, and radiotherapy plays a pivotal role in its therapeutic regimen. However, radiotherapy resistance is the main cause of therapeutic failure in patients. Our previous study revealed that Adiponectin Receptor 1 (AdipoR1) is involved in regulating radiation resistance in liver cancer patients treated with stereotactic body radiotherapy. To explore the mechanism, we performed high-throughput transcriptome sequencing of hepatocellular carcinoma cells with stable knockdown of AdipoR1. KEGG enrichment analysis indicated that the cell cycle and ubiquitination degradation pathways may be involved in the regulation of radiation resistance by AdipoR1.The knockdown of AdipoR1 can attenuate the radiation-induced G2/M phase arrest through cyclin B1.By the ubiquitination IP assay and a rescue experiment, we confirmed that CCNB1IP1 regulated the ubiquitination and degradation of cyclin B1. Combined with information from transcription factor database and AdipoR1 transcriptome sequencing, these results showed that estrogen receptor 1 (ESR1) may be a transcription factor of CCNB1IP1. We found that AdipoR1 promoted the translocation of ESR1 from the cytoplasm to the nucleus, and ESR1 inhibited the transcription of CCNB1IP1.Therefore, we propose that AdipoR1 regulates the ubiquitination level, cell cycle progression, and radiation resistance of HCC cells through the “AdipoR1 /ESR1/CCNB1IP1/cyclin B1” axis. This study will promote the development of novel targeted radiosensitizing drugs.

The origin and transmission of HIV-1 CRF80_0107 among two major first-tier cities in China

BackgroundCRF01_AE and CRF07_BC are the two most prevalent HIV-1 genotypes in China, and the co-circulation of these two genotypes has led to the continuous generation of CRF_0107 viruses in recent years. However, little is known about the origin and spread of CRF_0107 viruses thus far. This study focused on HIV-1 CRF80_0107, which we previously identified among the MSM population in Beijing and Hebei Province, to explore the demographic distribution, transmission links, and temporal-spatial evolutionary features of the HIV-1 CRF80_0107 strain in China.MethodsWith the partial pol region fragment of the HIV-1 CRF80_0107 subtype standard sequence as a reference, BLAST was used to search for highly similar sequences in the Los Alamos HIV Sequence Database, followed by preliminary subtype identification via COMET. Further phylogenetic and recombination breakpoint analyses were conducted to verify the subtypes and recombination patterns. We also performed a distance-based molecular network analysis to identify potential relationships among different HIV-positive individuals. In addition, spatiotemporal evolutionary dynamics analysis of the candidate CRF80_0107 sequences was performed via a Bayesian approach.ResultsA total of 36 partial pol gene sequences of HIV-1 CRF80_0107 were identified from 2009 to 2018 from 5 provinces in China. Phylogenetic and spatial-temporal dynamics analyses indicated that CRF80_0107 likely originated in Beijing around 2009 and spread to Guangdong Province around 2012. Population dynamics analysis revealed that CRF80_0107 experienced a significant increase in population size from 2009 to 2011 and then stabilized. The study also found that the number of cases in Guangdong Province was second only to that in Beijing and formed 2 relatively independent transmission clusters in the MSM population in Shenzhen, Guangdong Province.ConclusionsThe HIV-1 CRF80_0107 strain has spread to cities beyond its origin, particularly the MSM population in Shenzhen city, Guangdong Province, which is an area with a high incidence of HIV. This highlights the importance of continuous monitoring for the emergence and dynamic changes of novel HIV-1 recombinant viruses and the necessity of implementing effective preventive measures targeting specific populations in particular regions.

Wall teichoic acid glycosylation of bovine-associated Staphylococcus aureus strains.

Staphylococcus aureus (S. aureus) is one of the major causes of bovine mastitis, a disease with detrimental effects on health and wellbeing. Current control measures are costly, laborious and not always effective in eradicating S. aureus. The cell wall-linked polysaccharide wall teichoic acid (WTA) is highly immunogenic in humans and is considered as a prospective vaccine antigen based on promising pre-clinical studies in animals. WTA consist of polymerized ribitol-phosphate backbone that is modified with N-acetylglucosamine (GlcNAc) moieties in different configurations by the glycosyltransferases TarS (β-1,4-GlcNAc), TarM (α-1,4-GlcNAc) and TarP (β-1,3-GlcNAc). This study aimed to characterize the presence and genetic variation in tarS, tarM and tarP in bovine-associated S. aureus strains and how this impacts WTA-glycoprofile. Bioinformatic analyses of a whole genome sequence database consisting of 1047 S. aureus, 10 S. schweitzeri, and 6 S. argenteus strains showed that over 99% of strains contained tarS, 34 % also contained tarM, while 5 % of the strains encoded tarP in addition to tarS. The distribution of WTA-glycosyltransferase genes was similar to what has been reported for human-associated S. aureus strains. Phenotypic analysis of WTA glycosylation by flow cytometry corroborated with tarS/tarM/tarP gene presence. The WTA glycoprofile was variable between bovine-associated strains and the levels and ratios of GlcNAcylation were affected by growth conditions. Interestingly, a divergent tarM allele was present in strains of clonal complexes (CC) 49 and the mastitis-associated CC151, but its function was similar to canonical tarM. In conclusion, we demonstrated that bovine-associated S. aureus strains show similar variation in WTA GlcNAc decoration as human S. aureus strains, despite the presence of a divergent tarM allele.

Identification of a Novel HIV-1 Second-Generation Circulating Recombinant Form (CRF117_0107) in China.

Under the background of the main epidemic HIV strains (CRF01_AE and CRF07_BC) co-circulation in China, more HIV second-generation recombinant (SGR) strains with CRF01_AE and CRF07_BC as the backbone were also emerging. In this study, we characterize a novel HIV-1 second-generation circulating recombinant form (CRF117_0107) consisting of CRF01_AE and CRF07_BC fragments from three epidemiologically unrelated HIV-1-infected individuals. One near full-length genome (NFLG) sequence was amplified, sequenced, and spliced in two halves using RNA extracted from the plasma of a homosexual in Shenzhen, Guangdong Province. Two other NFLG sequences were obtained from the Los Alamos HIV Sequence Database under accession numbers KY201177 and MK397789, which were isolated from men who have sex with men (MSM) in Guangdong Province and Zhejiang Province, respectively. Phylogenetic analysis revealed that these NFLG sequences formed a monophyletic cluster with a high bootstrap value of 1.0. Recombination analysis demonstrated that the genome of CRF117_0107 was separated into three segments by two breakpoints. Further subregional phylogenetic analysis was performed that showed segment I+III (790-5990nt, 8295-9412nt) of CRF117_0107 originated from the CRF07_BC cluster, and Segment I+III (5991-8294nt) originated from the CRF01_AE cluster. The appearance of CRF117_0107 further highlights that HIV-1 SGR strains containing CRF01_AE and CRF07_BC will be generated more frequently and will most likely be more conducive to accelerating the spread of HIV in China. This study suggested it's essential to monitor HIV-1 second-generation CRFs among high-risk populations such as MSM for the epidemic and evolution dynamics of HIV-1 in China.

Advancing the Taxonomy of the Diatom Pseudo-nitzschia Through an Integrative Study Conducted in the Central and Southeastern Adriatic Sea.

The marine diatom genus Pseudo-nitzschia comprises cosmopolitan phytoplankton species commonly present in the Adriatic Sea. Species within the genus Pseudo-nitzschia have been of significant concern because they produce domoic acid (DA), which can cause amnesic shellfish poisoning (ASP). In this study, we identified Pseudo-nitzschia species along the Central and Southeastern Adriatic Sea, where monthly sampling carried out from February 2022 to February 2024 allowed for comprehensive species documentation. Pseudo-nitzschia species cell cultures isolated from the study areas were morphologically and molecularly analysed. Morphological analyses were performed using a scanning electron microscope (FE-SEM/STEM), while molecular analyses were conducted, targeting the ITS1-5.8S-ITS2, LSU, and rbcL regions, to confirm species identity. This integrative approach led to the identification of eight species: Pseudo-nitzschia allochrona, Pseudo-nitzschia calliantha, Pseudo-nitzschia delicatissima, Pseudo-nitzschia fraudulenta, Pseudo-nitzschia mannii, Pseudo-nitzschia multistriata, Pseudo-nitzschia pseudodelicatissima, and Pseudo-nitzschia subfraudulenta. Our findings underscore the value of a combined approach for reliable species identification and contribute to the development of genetic sequence databases that support the advancement of next-generation methods such as metabarcoding. This research emphasises the importance of combined morphological and molecular methods for the differentiation of the cryptic and pseudo-cryptic Pseudo-nitzschia species.

Guidelines for Gene and Genome Assembly Nomenclature.

The rapid increase in the number of reference-quality genome assemblies presents significant new opportunities for genomic research. However, the absence of standardized naming conventions for genome assemblies and annotations across datasets creates substantial challenges. Inconsistent naming hinders the identification of correct assemblies, complicates the integration of bioinformatics pipelines, and makes it difficult to link assemblies across multiple resources. To address this, we developed a specification for standardizing the naming of reference genome assemblies, to improve consistency across datasets and facilitate interoperability. This specification was created with FAIR (Findable, Accessible, Interoperable, and Reusable) practices in mind, ensuring that reference assemblies are easier to locate, access, and reuse across research communities. Additionally, it has been designed to comply with primary genomic data repositories, including members of the International Nucleotide Sequence Database Collaboration (INSDC) consortium, ensuring compatibility with widely used databases. While initially tailored to the agricultural genomics community, the specification is adaptable for use across different taxa. Widespread adoption of this standardized nomenclature would streamline assembly management, better enable cross-species analyses, and improve the reproducibility of research. It would also enhance natural language processing applications that depend on consistent reference assembly names in genomic literature, promoting greater integration and automated analysis of genomic data. This is a good time to consider more consistent genomic data nomenclature as many research communities and data resources are now finding themselves juggling multiple datasets from multiple data providers.

Immunoinformatics-guided design of a multiepitope peptide vaccine targeting the receptor-binding domain of SARS-CoV-2 spike glycoprotein: insights from Indonesian samples.

The emergence of new variants of SARS-CoV-2, including Alpha, Beta, Gamma, Delta, Omicron variants, and XBB sub-variants, contributes to the number of coronavirus cases worldwide. SARS-CoV-2 is a positive RNA virus with a genome of 29.9 kb that encodes four structural proteins: spike glycoprotein (S), envelope glycoprotein (E), membrane glycoprotein (M), and nucleocapsid glycoprotein (N). These proteins are vital for viral activity, with the S protein facilitating attachment and membrane fusion through the receptor-binding domain (RBD) located in the S1 subunit. The RBD recognizes and binds to the human angiotensin-converting enzyme 2 (ACE-2) protein. An immunoinformatic-aided design of a peptide-based multiepitope vaccine candidate targeting the RBD glycoprotein is constructed from the SARS-CoV-2 sequence data base from various regions of Indonesia (Jakarta, West Java, and Bali). The results show that the RBD region of with accession ID EPI_ISL_15982641 from West Java had the highest antigenicity of 0.5904. This isolate is non-toxic and non-allergenic and shows a total of 18 LBL epitopes, 72 CTL epitopes, and 98 HTL epitopes. The epitope that has the best overall binding affinity was GCHNKCAY for MHC-I and GGCVFSYVGCHNKCAYWV for MHC-II which show a binding affinity of -13.6 and -15.5 (kcal/mol), respectively. Therefore, this study aims to design an epitope vaccine candidate based on samples from Indonesia that has good characteristics and may have the potential to stimulate an immune response against SARS-CoV-2.

Comparative genomic analysis of Fusarium oxysporum f. sp. lycopersici reveals telomeric duplications of a lineage-specific region carrying SIX8 and PSL1 and genome-wide expansion of Foxy transposable elements.

Fusarium oxysporum f. sp. lycopersici (Fol), the causal agent of tomato wilt disease, is a soil-borne, vascular-colonizing fungal pathogen that severely impacts tomato production in most growing regions worldwide. Despite the availability of over thirty Fol genome sequences in public databases, only one chromosome-scale assembly exists, comprising the low sequence-coverage Fol4287 reference genome generated using Sanger sequencing. Thus, genome structural variation and comparative genomics analyses of Fol remain largely unexplored. Here, using third generation Nanopore long-read sequencing in combination with high-throughput chromosome conformation capture (Hi-C) data, we have independently constructed a high-quality assembly and annotation of the Fol007 race 2 genome that consists of 15 pseudochromosomes with 25 telomeres, including 10 telomere-to-telomere chromosomes. The Fol007 genome is predicted to contain 29,148 protein-encoding genes, including all known SIX genes except for SIX4, which is absent in Fol race 2. Compared to the genome of Fusarium verticillioides, the Fol007 genome contains four complete lineage-specific (LS) chromosomes, including chromosome 5 (Chr5), chromosome 13 (Chr13), and two small chromosomes, Chr14 and Chr15. Comparative genomic analysis between the newly assembled Fol007 genome and Fol4287_2010 deposited in the GenBank database reveals that the completely assembled Chr13 of Fol007 carrying all known SIX genes (except SIX4 and SIX8) and three novel candidate effector genes is relatively stable and corresponds to pathogenicity chromosome Chr14 in Fol4287_2010. It also reveals a telomeric duplication of an LS region carrying SIX8 and PSL1 on core chromosomes Chr6, Chr7 and Chr11, and LS chromosome Chr15, as well as the absence of segmental duplications on LS chromosomes of the Fol007 genome. Furthermore, compared to the genomes of Fol race 1 and non-pathogenic Fo strains, an active and specific Foxy transposable element, responsible for the inactivation of a second copy of SIX13, was identified and found to have expanded in the genomes of Fol race 2 and 3 strains. This element may contribute to Fusarium oxysporum genome evolution and has potential as a genetic marker for studying phylogenetic relationships among formae speciales of Fusarium oxysporum. These findings provide a basis for further genetic and genomic understanding of Fol evolution and virulence mechanisms employed by Fol and contribute another reference genome for Fusarium study more broadly.

A Clinical Metaproteomics Workflow Implemented within Galaxy Bioinformatics Platform to Analyze Host-Microbiome Interactions Underlying Human Disease.

Clinical metaproteomics reveals host-microbiome interactions underlying diseases. However, challenges to this approach exist. In particular, the characterization of microbial proteins present in low abundance relative to host proteins is difficult. Other significant challenges are attributed to using very large protein sequence databases, which impedes sensitivity and accuracy during peptide and protein identification from mass spectrometry data in addition to retrieving taxonomy and functional annotations and performing statistical analysis. To address these problems, we present an integrated bioinformatics workflow for mass spectrometry-based metaproteomics that combines custom protein sequence database generation, peptide-spectrum match generation and verification, quantification, taxonomic and functional annotations, and statistical analysis. This workflow also offers characterization of human proteins (while prioritizing microbial proteins), thus offering insights into host-microbe dynamics in disease. The tools and workflow are deployed in the Galaxy ecosystem, enabling the development, optimization, and dissemination of these computational resources. We have applied this workflow for metaproteomic analysis of numerous clinical sample types, such as nasopharyngeal swabs and bronchoalveolar lavage fluid. Here, we demonstrate its utility via the analysis of residual fluid from cervical swabs. The complete workflow and accompanying training resources are accessible on the Galaxy Training Network to equip non-experts and experienced researchers with the necessary knowledge and tools to analyze their data.

BenchAMRking: a Galaxy-based platform for illustrating the major issues associated with current antimicrobial resistance (AMR) gene prediction workflows

BackgroundThe Joint Programming Initiative on Antimicrobial Resistance (JPIAMR) networks ‘Seq4AMR’ and ‘B2B2B AMR Dx’ were established to promote collaboration between microbial whole genome sequencing (WGS) and antimicrobial resistance (AMR) stakeholders. A key topic discussed was the frequent variability in results obtained between different microbial WGS-related AMR gene prediction workflows. Further, comparative benchmarking studies are difficult to perform due to differences in AMR gene prediction accuracy and a lack of agreement in the naming of AMR genes (semantic conformity) for the results obtained. To illustrate this problem, and as a capacity-building exercise to encourage stakeholder involvement, a comparative Galaxy-based BenchAMRking platform was developed and validated using datasets from bacterial species with PCR-verified AMR gene presence or absence information from abritAMR.ResultsThe Galaxy-based BenchAMRking platform (https://erasmusmc-bioinformatics.github.io/benchAMRking/) specifically focusses on the steps involved in identifying AMR genes from raw reads and sequence assemblies. The platform currently comprises four well-characterised and published workflows that have previously been used to identify AMR genes using WGS data from several different bacterial species. These four workflows, which include the ISO certified abritAMR workflow, make use of different computational tools (or tool versions), and interrogate different AMR gene sequence databases. By utilising their own data, users can investigate potential AMR gene-calling problems associated with their own in silico workflows/protocols, with a potential use case outlined in this publication.ConclusionsBenchAMRking is a Galaxy-based comparison platform where users can access, visualise, and explore some of the major discrepancies associated with AMR gene prediction from microbial WGS data.

A lossless reference-free sequence compression algorithm leveraging grammatical, statistical, and substitution rules.

Deoxyribonucleic acid (DNA) or ribonucleic acid (RNA) sequence compressors for novel species frequently face challenges when processing wide-scale raw, FASTA, or multi-FASTA structured data. For years, molecular sequence databases have favored the widely used general-purpose Gzip and Zstd compressors. The absence of sequence-specific characteristics in these encoders results in subpar performance, and their use depends on time-consuming parameter adjustments. To address these limitations, in this article, we propose a reference-free, lossless sequence compressor called GraSS (Grammatical, Statistical, and Substitution Rule-Based). GraSS compresses sequences more effectively by taking advantage of certain characteristics seen in DNA and RNA sequences. It supports various formats, including raw, FASTA, and multi-FASTA, commonly found in GenBank DNA and RNA files. We evaluate GraSS's performance using ten benchmark DNA sequences with reduced number of repeats, two highly repetitive RNA sequences, and fifteen raw DNA sequences. Test results indicate that the weighted average compression ratios (WACR) for DNA and RNA sequences are 4.5 and 19.6, respectively. Additionally, the entire DNA sequence corpus has a total compression time (TCT) of 246.8 seconds (s). These results demonstrate that the proposed compression method performs better than several advanced algorithms specifically designed to handle various levels of sequence redundancy. The decompression times, memory usage, and CPU usage are also very competitive. Contact: anirban@klyuniv.ac.in.

An intron-split microRNA mediates cleavage of the mRNA encoded by low phosphate root in Solanaceae

Main conclusionA microRNA with a non-canonical precursor structure harbours an intron in between its miRNA-5p and miRNA-3p relevant for its biogenesis, is conserved across Solanaceae, and targets the mRNA of low phosphate root.Hundreds of miRNAs have been identified in plants and great advances have been accomplished in the understanding of plant miRNA biogenesis, mechanisms and functions. Still, many miRNAs, particularly those with less conventional features, remain to be discovered. Likewise, additional layers of regulation from miRNA generation to action and turnover are still being revealed. The current study describes a microRNA not previously identified given its unusual intron-split stem-loop structure, that has been previously observed only within the monocot-specific miRNA444 family. It shows its conservation across a branch of Solanales including agriculturally relevant Solanaceae family, where its transcripts had already been predicted in several species within sequence databases. The miRNA is absent in Arabidopsis thaliana but present in Solanum lycopersicum, Nicotiana benthamiana, Petunia axillaris, and Ipomoea nil. It proves that at least two different pri-miRNA variants are produced from this miRNA gene, one spliced and the other one retaining the intron. It demonstrates the dual function of its intron in the miRNA biogenesis. On the one hand, its presence in the pri-miRNA positively influences mature miRNA accumulation, but on the other hand, it needs to be removed from the pri-miRNA for efficient mature miRNA production. Finally, it sets low phosphate root as one of its targets, a protein known to be involved in root growth regulation under phosphate starvation in other plant species.

Single-cell transcriptomics reveals the heterogeneity and function of mast cells in human ccRCC.

The role of mast cells (MCs) in clear cell renal carcinoma (ccRCC) is unclear, and comprehensive single-cell studies of ccRCC MCs have not yet been performed. To investigate the heterogeneity and effects of MCs in ccRCC, we studied single-cell transcriptomes from four ccRCC patients, integrating both single-cell sequencing and bulk tissue sequencing data from online sequencing databases, followed by validation via spatial transcriptomics and multiplex immunohistochemistry (mIHC). We identified four MC signature genes (TPSB2, TPSAB1, CPA3, and HPGDS). MC density was significantly greater in ccRCC tissues than in normal tissues, but MC activation characteristics were not significantly different between ccRCC and normal tissues. Activated and resting MCs were defined as having high and low expression of MC receptors and mediators, respectively, whereas proliferating MCs had high expression of proliferation-related genes. The overall percentage of activated MCs in ccRCC tissues did not change significantly but shifted toward a more activated subpopulation (VEGFA+ MCs), with a concomitant decrease in proliferative MCs (TNF+ MCs) and resting MCs. An analysis of the ratio of TNF+/VEGFA+ MCs in tumors revealed that MCs exerted antitumor effects on ccRCC. However, VEGFA+MC was produced in large quantities in ccRCC tissues and promoted tumor angiogenesis compared with adjacent normal tissues, which aroused our concern. In addition, MC signature genes were associated with a better prognosis in the KIRC patient cohort in the TCGA database, which is consistent with our findings. Furthermore, the highest level of IL1B expression was observed in macrophages in ccRCC samples, and spatial transcriptome analysis revealed the colocalization of VEGFA+ MCs with IL1B+ macrophages at the tumor-normal interface. In conclusion, this study revealed increased MC density in ccRCC. Although the proportion of activated MCs was not significantly altered in ccRCC tissues compared with normal tissues, this finding highlights a shift in the MC phenotype from CTSGhighMCs to more activated VEGFA+MCs, providing a potential therapeutic target for inhibiting ccRCC progression.

Regional molecular transmission characteristics of newly reported HIV-infected students aged ≥18 years in Nanjing City from 2016 to 2022

To analyze the transmission characteristics of newly reported HIV-infected students aged ≥18 years in Nanjing City from 2016 to 2022 and provide evidence for AIDS publicity and intervention among young students. The pol region sequences of newly reported HIV-infected students and non-student HIV-infected individuals in Nanjing City from 2016 to 2022 were collected, and the BLAST tool was used to search the published global non-Nanjing reported HIV infection sequences in the LANL HIV database. The basic molecular transmission network and regional molecular transmission network were constructed using the HIV-TRACE in a pairwise genetic distance threshold of 1.0%. 332 sequences of infected students aged≥18 years in Nanjing City, 1 904 sequences of non-student-infected individuals in Nanjing City and 1 698 non-Nanjing-infected individuals were obtained. Among the 332 infected students, the main route of infection was homosexual (96.39%), and the subtypes were CRF01_AE (37.95%), CRF07_BC (37.65%) and CRF105_0107 (10.24%). There were 890 sequences in the regional molecular transmission network, of which 21.80% were infected students in Nanjing City, 39.89% were non-student-infected individuals in Nanjing City, and 38.31% were non-Nanjing-infected individuals. In the CRF105_0107 transmission cluster, non-student-infected individuals from Nanjing accounted for 66.95% (81/121), while in the CRF07_BC transmission cluster, non-Nanjing-infected individuals accounted for 56.66% (200/353). There were 1 644 edges connected to infected students within the regional molecular transmission network, with local transmission accounting for 64.72% and regional transmission accounting for 35.28%. Regional transmission was mainly in Guangdong Province (19.83%) and other cities in Jiangsu Province (4.50%). The HIV-1 subtypes of newly reported HIV-infected students aged≥18 years in Nanjing City are mainly CRF01_AE, CRF07_BC and CRF105_0107, with local transmission as the main transmission characteristics. There is transmission between students and non-students.