DNA Sequence Analysis Research Articles

Flea-Borne Rickettsioses and Scrub Typhus in Patients with Suspected Arbovirus Infection in Bangkok, Thailand.

Background: In urban Thailand, arboviral infections dominate diagnoses of acute undifferentiated fevers (AUFs) owing to their well-defined epidemiology and characteristic clinical presentations. However, rickettsial diseases, also endemic in this setting, remain under-recognized owing to challenges in early detection. Objective: This study aimed to identify potential rickettsial infections among patients with AUF in Bangkok and vicinity utilizing leftover nucleic acid extracted from serum samples from patients initially suspected of but negative for arbovirus infections. Materials and Methods: A total of 609 nucleic acid samples were screened for rickettsial bacteria using real-time PCR, targeting the 17-kDa common antigen gene of Rickettsia spp. and the 47-kDa gene of Orientia tsutsugamushi. Results: Nine samples were positive for Rickettsia spp. and two were positive for O. tsutsugamushi. DNA sequence and phylogenetic analyses based on partial 17-kDa antigen and citrate synthase (gltA) genes identified the Rickettsia-positive samples as R. typhi in eight cases and R. felis in one case. Analysis of the 56-kDa type-specific antigen gene identified the two O. tsutsugamushi isolates as Gilliam-related genotypes. Although rickettsial diseases typically present with mild symptoms, two patients with R. typhi infection (murine typhus) developed respiratory distress syndrome, highlighting the potential for rare but serious complications. Conclusion: This study underscores the critical importance of differential diagnosis and prompt, effective intervention to prevent complications in suspected cases.

High endemic freshwater mussel (Bivalvia: Unionida) diversity in western Borneo, with description of three new species

Abstract The freshwater mussels (Bivalvia: Unionida) of the biodiversity hotspot Sundaland are experiencing severe anthropogenic threats, whilst their diversity and distribution remain poorly understood. Here, we present the first modern-day data on Unionida diversity and distribution across western Borneo. Mussels were surveyed and collected in the upper Kapuas and Pawan river basins in West Kalimantan, Indonesia, and the Sambas Besar, Sarawak and Batang Sadong river basins in Sarawak, Malaysia. DNA sequencing (COI + 16S + ND1 + 18S + 28S) and morphological analyses were conducted to delineate and identify species, and reconstruct phylogenetic relationships and population genetics. Specimens belonged to six native genera and nine species, of which seven are endemic to Borneo and three are new to science, i.e. Ctenodesma mawonae Zieritz et al., sp. nov., Sarawak basin, and Ctenodesma bersinara Zieritz et al., sp. nov., and Rectidens lauris Zieritz et al., sp. nov., both Pawan basin. The monotypic genera Caudiculatus and Discomya were phylogenetically highly divergent from other known Gonideinae taxa, potentially indicating a separate tribe (for Caudiculatus) and subfamily (for Discomya). In addition, we report new records of the non-native Sinanodonta pacifica Heude, 1878 in the Batang Sadong and Kapuas river basins.

Fusion Genes Landscape of Lung Cancer Patients From Inner Mongolia, China.

Lung cancer is the leading cause of cancer-related deaths globally. Gene fusion, a key driver of tumorigenesis, has led to the identification of numerous driver gene fusions for lung cancer diagnosis and treatment. However, previous studies focused on Western populations, leaving the possibility of unrecognized lung cancer-associated gene fusions specific to Inner Mongolia due to its unique genetic background and dietary habits. To address this, we conducted DNA sequencing analysis on tumor and adjacent nontumor tissues from 1200 individuals with lung cancer in Inner Mongolia. Our analysis established a comprehensive fusion gene landscape specific to lung cancer in Inner Mongolia, shedding light on potential region-specific molecular mechanisms underlying the disease. Compared to Western cohorts, we observed a higher occurrence of ALK and RET fusions in Inner Mongolian patients. Additionally, we discovered eight novel fusion genes in three patients: SLC34A2-EPHB1, CCT6P3-GSTP1, BARHL2-APC, HRAS-MELK, FAM134B-ERBB2, ABCB1-GIPC1, GPR98-ALK, and FAM134B-SALL1. These previously unreported fusion genes suggest potential regional specificity. Furthermore, we characterized the fusion genes' structures based on breakpoints and described their impact on major functional gene domains. Importantly, the identified novel fusion genes exhibited significant clinical and pathological relevance. Notably, patients with SLC34A2-EPHB1, CCT6P3-GSTP1, and BARHL2-APC fusions showed sensitivity to the combination of chemotherapy and immunotherapy. Patients with HRAS-MELK, FAM134B-ERBB2, and ABCB1-GIPC1 fusions showed sensitivity to chemotherapy. In summary, our study provides novel insights into the frequency, distribution, and characteristics of specific fusion genes, offering valuable guidance for the development of effective clinical treatments, particularly in Inner Mongolia.

Polymorphism of the BMP15 and GDF9 genes in Romanov, Wrzosówka, and Podhale Zackel sheep in Poland

Various mutations in the BMP15 and GDF9 genes belonging to the transforming growth factor-β (TGF-β) superfamily have been shown to affect the prolificacy in sheep. Increasing the litter size is critical to the profitability of sheep production, but focusing on increasing the number of lambs born is only one of many factors that make sheep farming profitable. The study aimed to identify polymorphisms in BMP15 and GDF9 in Romanov, Wrzosówka, and Podhale Zackiel sheep breeds. The polymorphism of the coding parts of the BMP15 and GDF9 genes was determined based on DNA sequence analysis. Statistical analysis showed a significant relationship (p≤0.05) between the identified leucine deletion at the BMP15 locus (c.28_30delCTT) and increased fertility in heterozygous sheep. Statistically significant differences in the level of fertility were also identified between individuals with a mutation in the GDF9 gene - (c.978A> G, G6 - c.994G> A) and carriers of mutations in two BMP15 genes and GDF9 simultaneously.

DNA Barcoding Analysis of Betok Fish (Anabas testudineus) from Kampar, Riau Based on Cytochrome Oxidase Subunit I (COI)

Ikan betok (Anabas testudineus, Bloch 1792) is one of fishes which is a member of family Anabantidae. Betok is an endemic fish in Riau Province. The scientific study about DNA barcoding of betok fish is still rarely. This research aims to analyze the DNA barcode sequence of cytochrome c oxidase I (COI) on betok fish. Methods were used in this study such as sampling, total DNA extraction, PCR, electrophoresis, sequencing and data analysis. The COI sequence studied had size of 694 bp. The BLASTn analysis showed that the betok fish had the highest similarity of 99.7% with A. testudineus and the lowest 93.00% with A. cobojinus. There was one nucleotide that characterize the betok fish based on the COI sequence namely nucleotide number 309. This study may enrich the DNA barcode database of betok fish in GeneBank.

Microbial populations vary between the upper and lower respiratory tract, but not within biogeographic regions of the lung of healthy horses

Understanding normal microbial populations within areas of the respiratory tract is essential, as variable regional conditions create different niches for microbial flora, and proliferation of commensal microbes likely contributes to clinical respiratory disease. The objective was to describe microbial population variability between respiratory tract locations in healthy horses. Samples were collected from four healthy adult horses by nasopharyngeal lavage (NPL), transtracheal aspirate (TTA), and bronchoalveolar lavage (BAL) of six distinct regions within the lung. Full-length 16S ribosomal DNA sequencing and microbial profiling analysis was performed. There was a large amount of diversity, with over 1797 ASVs identified, reduced to 94 taxa after tip agglomeration and prevalence filtering. Number of taxa and diversity were highly variable across horses, sample types, and BAL locations. Firmicutes, proteobacteria, and actinobacteria were the predominant phyla. There was a significant difference in richness (Chao1, p = 0.02) and phylogenetic diversity (FaithPD, p = 0.01) between NPL, TTA, and BAL. Sample type (p = 0.03) and horse (p = 0.005) contributed significantly to Bray-Curtis compositional diversity, while Weighted Unifrac metric was only affected by simplified sample type (NPL and TTA vs BAL, p = 0.04). There was no significant effect of BAL locations within the lung with alpha or beta diversity statistical tests. Overall findings support diverse microbial populations that were variable between upper and lower respiratory tract locations, but with no apparent difference in microbial populations of the six biogeographic regions of the lung, suggesting that BAL fluid obtained blindly by standard clinical techniques may be sufficient for future studies in healthy horses.

Genetic Analysis and Fine Mapping of a New Rice Mutant, Leaf Tip Senescence 2.

Premature leaf senescence significantly reduces rice yields. Despite identifying numerous factors influencing these processes, the intricate genetic regulatory networks governing leaf senescence demand further exploration. We report the characterization of a stably inherited, ethyl methanesulfonate(EMS)-induced rice mutant with wilted leaf tips from seedling till harvesting, designated lts2. This mutant exhibits dwarfism and early senescence at the leaf tips and margins from the seedling stage when compared to the wild type. Furthermore, lts2 displays a substantial decline in both photosynthetic activity and chlorophyll content. Transmission electron microscopy revealed the presence of numerous osmiophilic granules in chloroplast cells near the senescent leaf tips, indicative of advanced cellular senescence. There was also a significant accumulation of H2O2, alongside the up-regulation of senescence-associated genes within the leaf tissues. Genetic mapping situated lts2 between SSR markers Q1 and L12, covering a physical distance of approximately 212 kb in chr.1. No similar genes controlling a premature senescence leaf phenotype have been identified in the region, and subsequent DNA and bulk segregant analysis (BSA) sequencing analyses only identified a single nucleotide substitution (C-T) in the exon of LOC_Os01g35860. These findings position the lts2 mutant as a valuable genetic model for elucidating chlorophyll metabolism and for further functional analysis of the gene in rice.

Land use intensity is a major driver of soil microbial and carbon cycling across an agricultural landscape

Soil carbon (C) storage is a critical ecosystem function that underpins human health and well-being. The acceleration of human-driven land use change, such as agricultural intensification, is a major driver of soil C loss globally. Developing sustainable land use practices that enhance agricultural productivity whilst protecting essential ecosystem functions such as soil C storage is vital. The soil microbiome has a critical role in regulating soil biogeochemical cycling processes, including soil C cycling. Examining the impacts of land use intensity on the soil microbiome enables us to assess the potential effects on long-term soil C stocks. Using metagenomic DNA sequencing and phospholipid fatty acid analysis, we investigated differences in the activity, diversity, and function of the soil microbiome associated with five contrasting land uses across an agricultural landscape. The land uses covered a gradient of disturbance intensities and included remnant native forest, regenerating native bush, exotic plantation forest, dryland pasture, and irrigated pasture. We identified pronounced differences in the soil microbiome associated with each land use, including the diversity and abundance of microbial C and nitrogen (N) cycling genes. Notably, intensive agricultural land uses had a significantly higher diversity and abundance of microbial C-degrading genes, whilst land uses of remnant native forest had the lowest diversity and abundance of microbial C-degrading genes. Our findings suggest that intensive agricultural land use may increase the functional potential of the soil microbiome to mineralize soil C, potentially resulting in a greater loss of soil C as respired CO2 into the atmosphere. This research may be used to support the development of sustainable management practices that promote the persistence of soil C across agricultural landscapes, such as the protection of remnant native forest fragments and greater incorporation of regenerating native vegetation.

BRCA1 foci test as a predictive biomarker of olaparib response in ovarian cancer patient-derived xenograft models.

Standard therapy for high-grade ovarian carcinoma includes surgery followed by platinum-based chemotherapy and poly-ADP ribose polymerase inhibitors (PARPis). Deficiency in homologous recombination repair (HRD) characterizes almost half of high-grade ovarian carcinomas and is due to genetic and epigenetic alterations in genes involved in HR repair, mainly BRCA1/BRCA2, and predicts response to PARPi. The academic and commercial tests set up to define the HRD status of the tumor rely on DNA sequencing analysis, while functional tests such as the RAD51 foci assay are currently under study, but have not been validated yet and are available for patients. In a well-characterized ovarian carcinoma patient-derived xenograft platform whose response to cisplatin and olaparib, a PARPi, is known, we assessed the association between the BRCA1 foci score, determined in formalin-fixed paraffin-embedded tumor slices with an immunofluorescence technique, and other HRD biomarkers and explored the potential of the BRCA1 foci test to predict tumors' response to cisplatin and olaparib. The BRCA1 foci score was associated with both tumors' HRD status and RAD51 foci score. A low BRCA1 foci score predicted response to olaparib and cisplatin, while a high score was associated with resistance to therapy. As we recently published that a low RAD51 foci score predicted olaparib sensitivity in our xenobank, we combined the two scores and showed that the predictive value was better than with the single tests. This study reports for the first time the capacity of the BRCA1 foci test to identify HRD ovarian carcinomas and possibly predict response to olaparib.

Neurigona susanrocesae, the first Neurigona species from the Philippines

The first species of the genus Neurigona is described from the Philippines. The diagnosis, description and molecular data based on the partial DNA sequence of mitochondrial cytochrome oxidase subunit I gene were documented. The Neurigona species exhibits distinct characteristics, including a sub-apical dorsal stylus resembling an arista, a peduncle on the seventh abdominal segment, a flat dorsal postcranium, elongated legs devoid of prominent setae or bristles, and a distinctive rectangular bend in wing vein M. Based on the DNA sequence analysis, the unknown species is 90.06% similar to Cyrtona sp. and Neurigona zhejiangensis. However, 10% of its DNA sequences are significantly variable making it a totally different species. The specimen was verified by a dipterist as a species belonging to Genus Neurigona under Family Dolichopodidae. These flies are recognized for their lengthy legs and sizable yellow bodies. The species was named as N. susanrocesae as a tribute to the late Susan Roces, the “Queen of Philippine Movies”.

Piecing together the taxonomic puzzle: Generic delimitation in the Stoebe clade of Cape daisies (Asteraceae: Gnaphalieae) and a synopsis of the species

The Stoebe clade is a Cape-centred lineage comprising ca. 70 species, currently in eight genera. Despite recent generic taxonomic changes, many generic characters are ambiguous, and generic circumscription is likely to remain unstable in the absence of a robust species-level phylogeny. Here, we present a hypothesis of species relationships in the clade, based on parsimony and Bayesian analyses of DNA sequence and morphological characters, in order to assess generic monophyly. The trees based on morphology alone and plastid sequences alone are poorly resolved, with few well-supported nodes, while those based on nuclear sequences alone are better resolved. There is no evidence of phylogenetic conflict, and concatenation of all three data sources produces a well-resolved tree with many highly-supported nodes. We show that none of the currently-accepted genera are monophyletic, with the exception of Muscosomorphe, which is supported as monospecific. Amphiglossa species are recovered in two separate clades, requiring the resurrection of the genus Pterothrix to house ten of the twelve species. All the characters used to circumscribe the remaining genera, such as the possession of ray florets, are shown to be homoplasious. As a result, these remaining genera (Dicerothamnus, Disparago, Elytropappus, Myrovernix, Seriphium, and Stoebe) are here synonymised with Stoebe. The main morphological synapomorphy for this enlarged concept of Stoebe is the aggregation of the tiny capitula into conflorescences with varying degrees of complexity and congestion. The proposed taxonomy for the Stoebe clade comprises just four genera: Amphiglossa, Muscosomorphe, Pterothrix, and Stoebe. We present the new combinations, keys and generic descriptions, as well as keys to the species.

Occurrence, distribution, and genetic diversity of faba bean viruses in China.

With worldwide cultivation, the faba bean (Vicia faba L.) stands as one of the most vital cool-season legume crops, serving as a major component of food security. China leads global faba bean production in terms of both total planting area and yield, with major production hubs in Yunnan, Sichuan, Jiangsu, and Gansu provinces. The faba bean viruses have caused serious yield losses in these production areas, but previous researches have not comprehensively investigated this issue. In this study, we collected 287 faba bean samples over three consecutive years from eight provinces/municipalities of China. We employed small RNA sequencing, RT-PCR, DNA sequencing, and phylogenetic analysis to detect the presence of viruses and examine their incidence, distribution, and genetic diversity. We identified a total of nine distinct viruses: bean yellow mosaic virus (BYMV, Potyvirus), milk vetch dwarf virus (MDV, Nanovirus), vicia cryptic virus (VCV, Alphapartitivirus), bean common mosaic virus (BCMV, Potyvirus), beet western yellows virus (BWYV, Polerovirus), broad bean wilt virus (BBWV, Fabavirus), soybean mosaic virus (SMV, Potyvirus), pea seed-borne mosaic virus (PSbMV, Potyvirus), and cucumber mosaic virus (CMV, Cucumovirus). BYMV was the predominant virus found during our sampling, followed by MDV and VCV. This study marks the first reported detection of BCMV in Chinese faba bean fields. Except for several isolates from Gansu and Yunnan provinces, our sequence analysis revealed that the majority of BYMV isolates contain highly conserved nucleotide sequences of coat protein (CP). Amino acid sequence alignment indicates that there is a conserved NAG motif at the N-terminal region of BYMV CP, which is considered important for aphid transmission. Our findings not only highlight the presence and diversity of pathogenic viruses in Chinese faba bean production, but also provide target pathogens for future antiviral resource screening and a basis for antiviral breeding.

Discovery of Botryosphaeria eucalypti sp. nov. from blighted Eucalyptus leaves in India

Eucalyptus spp. are undoubtedly one of the most favored plantation trees globally. Accurately identifying Eucalyptus pathogens is therefore crucial for timely disease prevention and control. Recently, symptoms of a leaf blight disease were observed on Eucalyptus trees in plantations at Jhajjar and Karnal in the state of Haryana, northern India. Asexual morphs resembling the features of the Botryosphaeriaceae were consistently isolated from the symptomatic leaves. Morphological features coupled with DNA sequence analysis confirmed a novel species, which is described and illustrated here as Botryosphaeria eucalypti sp. nov. Conidia of the new taxon are longer and wider than those of its phylogenetic neighbors. A distinct phylogenetic position for the new taxon was established through combined analysis of the internal transcribed spacer (ITS), partial translation elongation factor-1α (tef1) and partial β-tubulin (tub2) regions. Recombination analysis provided additional support for the new species hypothesis. The pathogenicity of the novel species was proved on Eucalyptus leaves, and Koch's postulates were fulfilled. The discovery of new Botryosphaeria species is important because it will help in understanding the species diversity, host range, possible threats and disease control in the long run.

Proof of ssDNA degraded from dsDNA for ET recombination

The widely used ET recombination requires an ssDNA product degraded by Rac phage protein E588 from dsDNA for strand invasion. However, proof of the ssDNA product is still elusive. The study provided three levels of proof sequentially. The probable ssDNAs degraded by E588 from the fluorescent plus-, minus-, or double-stranded dsDNA pET28a-xylanase exhibited a half fluorescence intensity of the corresponding dsDNAs, equivalent to the E588 degradation nucleotides half that of the total nucleotides degraded from the corresponding dsDNA. The ssDNA product degraded by E588 from the fluorescent minus-stranded dsDNA was confirmed by gradient gel-electrophoresis and two nuclease degradation reactions. Degraded by E588 from the dsDNA pET28a-xylanase that had a phosphorothioated plus-stranded 5′-terminus, the plus-stranded ssDNA product was separated via gel electrophoresis and recovered via a DNAclean kit. The recovered ssDNA product was proven to have intact 5′- and 3′-ends by DNA sequencing analysis. This study provides a solid foundation for the mechanism of ssDNA invasion.

MDB-16. CLINICAL AND MOLECULAR CHARACTERISTICS OF JAPANESE MEDULLOBLASTOMA, WNT-ACTIVATED

Abstract BACKGROUND Medulloblastomas are at least classified into four core subgroups, including WNT-activated, SHH-activated, group 3 and group 4. These molecular subgroups have been introduced in the latest WHO classification, which has become fundamental and indispensable. Medulloblastoma, WNT-activated (WNT-MB) accounts for about 10% of all medulloblastomas, and their prognosis is excellent, and it is desired to ensure that patients with WNT-MBs are diagnosed exactly. In this study, we have summarized results of molecular analysis of Japanese MBs collected by Japanese Pediatric Molecular Neuro-oncology Group (JPMNG) study, and analyzed clinical and molecular characteristics of WNT-MBs. METHODS We enrolled 267 cases of medulloblastoma from 44 centers participating in the JPMNG study since 2012.Central histopathological diagnosis was performed by three neuropathologists, and molecular genetic diagnosis was performed using molecular subgrouping, DNA sequencing (CTNNB1), and copy number analysis. RESULTS A total of 244 cases were finally analyzed, and the 4-subgroup classification was WNT: 39 (16%), SHH: 61 (25%), group3: 42 (17%), and group4: 102 (41.8%), respectively. The age at diagnosis was 3 years or older (6-40 years) in all cases, with no cases occurring in infants, the majority being children (6-18 years) (31/39: 79.5%), and 8 cases (8/39: 20.5%) in adults older than 18 years. Histological features were classic (CMB) in the majority of cases and large cell (LCA) type in only 2 cases. The positivity rate of nuclear β-catenin by Immunological staining was 9 out of 12 (75%) of the cases examined. While, CTNNB1mutation was positive in 39 of 39 cases (100%) and monosomy 6 was also positive in 35 of 35 cases (100%), respectively. The 5-year survival rate was 91.9%. CONCLUSIONS Clinical characteristics of Japanese WBT-MB were almost similar to those reported in other countries, which showed excellent prognosis, and their molecular genetic status were also identical with results from other ethnics.

Ophiostoma haidaense, sp. nov., a new member of the O. piceae species complex associated with yellow-cedar, Callitropsis nootkatensis

ABSTRACT Ophiostoma haidanensis is described as a new species of the Ophiostoma piceae complex isolated from yellow-cedar (Callitropsis nootkatensis (D. Don) Oerst. ex D.P. Little) sapwood in the Haida Gwaii island archipelago and the North Coast of British Columbia, Canada. The fungus is characterized by the production of a typical sporothrix-like asexual morph but is distinguished morphologically from other members of the O. piceae species complex by its large, multiseptate primary conidia. Phylogenetic analysis of DNA sequences from the nuc rDNA internal transcribed spacer region ITS1-5.8S-ITS2 (ITS) and the β-tubulin (BTUB) and translation elongation factor 1-α (TEF1) genes supports the inclusion of O. haidensis as a distinct member within the O. piceae complex. To our knowledge, this is the first report of a blue stain fungus infecting yellow-cedar, an ecologically, culturally, and economically important conifer naturally distributed along the coastal forests of the Pacific Northwest in North America.

Single-Gene Phylogeny of Brucella Melitensis in Milk Samples of Ewes in Kalar District, Iraqi Kurdistan Region

The present study aimed to investigate the frequency rate of ovine brucellosis in Kalar district, Iraqi Kurdistan Region and to identify Brucella species in the milk of seropositive ewes via DNA sequencing and phylogenetic analyses. Blood samples were collected from 300 ewes and subjected to the Rose Bengal Plate Test (RBPT), and milk samples of the seropositive animals were collected and underwent a PCR assay using genus- and species-specific primers. Eventually, the positive PCR products were processed for nucleotide sequencing and single-gene phylogenetic analyses. The RBPT revealed a high percentage (11.33%) of seropositive ewes (34 out of 300), and the PCR assay revealed 35.3% (12/34) Brucella genus-positive DNA templates. At the species level, 58.3% (7/12) of templates were positive for the melitensis-specific primers and negative for the abortus and ovis-specific primers, whereas the other five were negative for all species-specific primers used in this study (abortus, melitensis and ovis). The nucleotides sequence of Brucella strains detected in this study showed a 99-100% homology with eight GenBank-published B. melitensis strains, and the single gene phylogenetic analysis showed a close relationship between these strains and the B. melitensis biovar 3 strain NI, China. The high percentage of seropositive ewes showed that brucellosis represents a major obstacle to the animals’ production improvement and a major risk to public health in the Iraqi Kurdistan Region and the single gene phylogenetic analysis confirms its transboundary spread and attracts attention for more strict control programs.

Ribbon worms (phylum Nemertea) from Bodega Bay, California: A largely undescribed diversity.

The diversity of nemerteans along the Pacific coast of the United States is regarded as well characterized, but there remain many cryptic, undescribed, and "orphan" species (those known only in their larval form). Recent sampling of nemerteans in Oregon and Washington has begun to fill in these taxonomic gaps, but nemertean diversity in California has received relatively little attention over the past 60 years. During the summers of 2019 and 2020, nemertean specimens were collected from 20 locations in the Bodega Bay region of northern California, USA, including rocky intertidal shores, sandy beaches, mudflats, and other habitats. Based on morphological assessment and DNA sequence analysis (partial Cytochrome Oxidase I and 16S rRNA genes), our surveys identified 34 nemertean species. Only 13 of these (38%) can be confidently assigned to described species. Another 11 represent species that are new to science, including members of the genera Riserius, Nipponnemertes, Poseidonemertes, Zygonemertes, Nemertellina, Oerstedia, and three species of uncertain affiliation. The remaining ten species include undescribed or cryptic species of uncertain status that have been found previously along the Pacific Coast of North America. Our surveys also document extensions of known geographic ranges for multiple species, including the first records in California of Antarctonemertesphyllospadicola, Cephalothrixhermaphroditica, and Maculauraoregonensis. This is the first report of the genus Nemertellina in the northeast Pacific and Riserius in California. Overall, our findings highlight how much remains to be learned about the diversity and distribution of nemerteans in the northeast Pacific.

Molecular characteristics of dedifferentiated liposarcoma with broad rhabdomyosarcomatous differentiation.

e23530 Background: Dedifferentiated liposarcoma (DDLPS) with broad rhabdomyosarcomatous differentiation has a worse prognosis compared with other types of DDLPS. Therefore, it is of importance to recognize its molecular characteristics in order to investigate individualized treatment strategies. Methods: We conduct next-generation sequencing (NGS) including DNA and RNA sequencing analysis in 5 DDLPS patients with broad rhabdomyosarcomatous differentiation and perform comparative analysis between well-differentiated and dedifferentiated components from the patients. Results: In the ten samples, several somatic copy-number variations (CNVs) including the gain of 12q are considered as frequent molecular alterations. IGFN1, FGFR4, BLM, RICTOR, RUNX1 and TOP1 mutation is the high-frequency single-nucleotide variations (SNVs). Novel BEST3/CD63/LGR5/CENPL-MDM2 and ERBB3-TAFA2 fusion genes are identified in this subtype of tumor. The comparative analysis between well-differentiated and dedifferentiated components recognized two categories of molecular variations: shared variations, such as FGFR4 and BLM mutation, ERBB3-TAFA2 fusion, and MYC amplification that are associated with tumorigenesis, and dedifferentiated-specific variations including GRIN2A, FOXP1 and TP53 mutation, CERS4-PTPRB and TRIO-TERT fusion, and ESR1 amplification, which are related with malignant progression. After surgery, the patient received adjuvant therapy. The mean follow-up of the patients was 30 months, and the mean recurrence-free survival (RFS) was 9.6 months. Conclusions: This NGS analysis disclosed the possible progression mechanisms and therapeutic targets in DDLPS with broad rhabdomyosarcomatous differentiation and provided insights contributing to the refinement of management in this tumor. [Table: see text]

Phytophthora polonica and Phytophthora hydropathica from Clade 9 Associated with Alder Decline in Bulgaria.

A number of Phytophthora species have been identified as destructive plant pathogens and invasive species. They have the potential to affect a wide range of host plants and cause diseases in agricultural and forest ecosystems. Two Phytophthora species from rhizosphere soil samples collected from declining Alnus glutinosa in Bulgaria were isolated in the autumn of 2022. They were identified as Phytophthora polonica and Phytophthora hydropathica according to the DNA sequence analysis of the ITS region, as well as their morphological and physiological characteristics. The pathogenicity of both species to common and gray alder was evaluated by the inoculation of detached leaves and cuttings. Experimental data proved that P. polonica and P. hydropathica are able to cause leaf necrosis not only on A. glutinosa from which they were derived, but also on A. incana. No significant deference in the aggressiveness of the studied isolates from both Phytophthora species against the two tested plants was observed. Therefore, P. polonica and P. hydropathica were determined as potential threats for alder ecosystems in the country. This is the first report for the isolation of P. polonica in Bulgaria and represents the most southeastern point of the species distribution in Europe.