Molecular Evidence Research Articles

Two new species and three new records of Diachea (Physarales) from China based on morphological and molecular evidence

Diachea is an important genus of myxomycetes, recognized for its ecological role and wide distribution. This study aimed to expand knowledge of species diversity within this genus in China. We collected Diachea specimens from various locations in Shaanxi and Sichuan provinces and characterized them through morphological analysis and phylogenetic analysis using four genetic markers: small subunit ribosomal RNA (nSSU), translation elongation factor 1-alpha (EF-1α), mitochondrial small subunit (mtSSU), and alpha-tubulin gene (α-Tub). Based on these analyses, we describe two new species, namely, Diachea plectophylla and D. sichuanensis, discovered in the Shaanxi and Sichuan provinces, respectively. Diachea plectophylla is distinguished by its dense, rigid capillitium, spore warts, and distinct separation of capillitium ends from the peridium. Diachea sichuanensis, closely related to D. leucopodia, is identified by its blunt-headed columella, clustered spore warts, and robust stalks. In addition to these new species, we recorded five previously documented species, including D. bulbillosa in Gansu province, D. leucopodia in Yunnan and Sichuan provinces, and D. subsessilis in Sichuan province. Detailed descriptions, micrographs, taxonomic comparisons, and an identification key are provided to aid in accurate identification. The discovery of these new species not only enhances the known diversity of slime molds in the region but also provides valuable information for future studies on their geographical distribution and ecological relationships.

Taxonomic notes on Sophora phulangkaensis, a new species of Sophora (Fabaceae – Sophoreae) from Thailand based on morphological and molecular evidence

Sophora phulangkaensis is illustrated and newly described here. The species shows similarities with its congener, S. exigua, by having an obovate terminal leaf let and up to 15 leaflets but differs in the floralparts. A morphological comparison between the two species is provided along with a discussion. The phylogenetic place- ment of then ewspecies is presented and discussed based on molecular evidence by nuclear (ITS) and chloroplast markers (matK). The results suggest that the new species is nested into sect. Rubriflorae, which includes S. exigua and S. huamotensis. The circumscription of sect. Rubriflorae is expanded after being redefined by our findings. In addition, the updated key to species of the genus Sophora for the Flora of Thailand is given based on recent specimen observations. Additionally, lectotypes for S. exigua, S. tonkinensis, and S. violacea var. pilosa are designated.

A new species of Autocrates (Coleoptera: Trictenotomidae) revealed by DNA barcoding and morphological evidence

Autocrates soni sp. nov. is described from Vietnam based on morphological and molecular evidence. This beetle is characterised by reddish-brown pubescence on its ventral side and much shorter setae on its femora compared to the closely related species Autocrates vitalisi Vuillet, 1912. The validity of this new species is strongly supported by DNA barcode data from all known species of the genus Autocrates.

Phylogenetic and taxonomic revisions of Jurassic sea stars support a delayed evolutionary origin of the Asteriidae.

The superorder Forcipulatacea is a major clade of sea stars with approximately 400 extant species across three orders (Forcipulatida, Brisingida, Zorocallida). Over the past century, the systematics of Forcipulatacea have undergone multiple revisions by various authors, with some considering numerous families such as Asteriidae, Zoroasteridae, Pedicellasteridae, Stichasteridae, Heliasteridae, Labidiasteridae, and Neomorphasteridae, while others recognized only two families (i.e., Asteriidae and Zoroasteridae). Recent molecular analyses have shown the artificial nature of some of these groupings. Notably, four well-supported clades (Zorocallida, Brisingida, Stichasteridae, and Asteriidae) emerged from a synthesis of morphological and molecular evidence. The majority of extinct forcipulatacean species have been placed in modern families. However, many of these fossil species are in need of revision, especially those species placed within the Asteriidae, the largest of all forcipulatacean families. In light of recent advancements in forcipulatacean systematics, we comprehensively reassess six well-preserved Jurassic forcipulatacean taxa, including the earliest crown-group members from the Hettangian (∼201.4 Ma), and also describe two new Jurassic genera, Forbesasterias gen. nov. and Marbleaster gen. nov. We assembled the largest and most comprehensive phylogenetic matrix for this group, sampling 42 fossil and extant forcipulatacean species for 120 morphological characters. To infer phylogenetic relationships and construct an evolutionary timeline for the diversification of major clades, we conducted a Bayesian tip-dating analysis incorporating the fossilized birth-death process. A total of 13 fossil species were sampled in our analysis, including six taxonomically revaluated herein, two recently reappraised species from the Jurassic, and five additional species from the Cretaceous and Miocene. Contrary to prior assumptions, our results indicate that none of the Jurassic taxa investigated belong to Asteriidae or any other modern families, and instead represent stem-forcipulatids. Furthermore, our phylogenetic results suggest that Asteriidae likely originated during the late Cretaceous. Our findings highlight a greater early diversity within the Forcipulatacea than previously presumed, challenging existing perceptions of the evolutionary history of this significant clade of marine invertebrates.

Morphological and molecular identification for four new wood-inhabiting species of Lyomyces (Basidiomycota) from China

Fungi are one of the most diverse groups of organisms on Earth, in which the wood-inhabiting fungi play an important role in forest ecosystem processes and functions. Four new wood-inhabiting fungi, Lyomyces hengduanensis, L. niveomarginatus, L. wumengshanensis and L. zhaotongensis, are proposed, based on morphological features and molecular evidence. Lyomyces hengduanensis differs in the brittle basidiomata with pruinose hymenial surface, a monomitic hyphal system and ellipsoid basidiospores (3.5–6 × 3–4.5 µm). Lyomyces niveomarginatus is distinguished by the subceraceous basidiomata with crackled hymenial surface, a monomitic hyphal system and ellipsoid basidiospores (4.5–7 × 3–4 µm). Lyomyces wumengshanensis is distinguished by the grandinioid hymenial surface, a monomitic hyphal system and ellipsoid to broad ellipsoid basidiospores (4–6 × 3–5 µm). Lyomyces zhaotongensis is unique in the grandinioid hymenial surface, a monomitic hyphal system and broadly ellipsoid basidiospores measuring as 2.6–3.5 × 2.5–3 µm. Sequences of ITS and nLSU rRNA markers of the studied samples were generated and phylogenetic analyses were performed using the Maximum Likelihood, Maximum Parsimony and Bayesian Inference methods. The phylogram, based on the ITS+nLSU rDNA gene regions, included three genera within the Schizoporaceae viz. Fasciodontia, Lyomyces and Xylodon, in which the four new species were grouped into Lyomyces. The phylogenetic tree inferred from the ITS sequences highlighted that L. hengduanensis group with L. zhaotongensis and then closely grouped with L. crustosus, L. ochraceoalbus, and L. vietnamensis. The new taxon L. niveomarginatus was retrieved as a sister to L. juniperi. The new species L. wumengshanensis was sister to L. macrosporus. The new taxon L. zhaotongensis grouped with L. hengduanensis and then closely grouped with L. crustosus, L. ochraceoalbus and L. vietnamensis.

First molecular evidence of bovine hemoplasmas in houseflies (Musca domestica)

BackgroundHemoplasma infections in cattle are caused by Mycoplasma wenyonii and Candidatus Mycoplasma haemobos and induce asymptomatic or chronic infections but occasionally lead to life-threatening hemolytic anemia. Despite the global distribution of bovine hemoplasmas, information regarding their transmission vectors and prevalence is still lacking in the Republic of Korea. Therefore, this study aims to investigate the infection rate of bovine hemoplasma in cattle and houseflies and to assess the risk factors associated with hemoplasma infection in cattle.MethodsOverall, 376 blood samples were collected from Korean indigenous cattle (male, 10−13 months old), along with 2,690 houseflies (Musca domestica) from the same farm where the cattle were raised. PCR assays targeting the 16S rRNA gene were performed to detect hemoplasmas, and positive samples were sequenced.ResultsThe infection rate of bovine hemoplasmas was 50.8% (191/376) in cattle and 7.4% in pooled houseflies. Among cattle, 18.6% (70/376) and 20.0% (75/376) tested positive for M. wenyonii and Candidatus M. haemobos, respectively. Conversely, in houseflies, Candidatus M. haemobos was more frequently detected (5.9%) than M. wenyonii (0.7%). Co-infection was 12.2% (46/376) in cattle and 0.7% in flies. Furthermore, hemoplasma infection was significantly associated with the grazing experience of their dams. Cattle born to cows with grazing experience exhibited a higher risk for M. wenyonii infection (odds ratio [OR] = 1.62; 95% confidence interval [CI]: 1.03−2.55; P = 0.045), whereas these cattle had a lower risk for Candidatus M. haemobos infection (OR = 0.32, 95% CI: 0.19−0.74; P = 0.000) than animals born to cows without grazing experience. The sequences obtained from houseflies were confirmed as Candidatus M. haemobos, which displayed high similarity (98.2−100%) to those from cattle obtained in this study.ConclusionsTo our knowledge, this study represents the first report of bovine hemoplasmas identified in houseflies. This molecular evidence suggests that houseflies may be possible vectors for Candidatus M. haemobos.

Molecular Evidence for the Axial Coordination Effect of Atomic Iodine on Fe‐N4 Sites in Oxygen Reduction Reaction

AbstractWe present a molecular‐scale investigation of the axial coordination effect of atomic iodine on Fe‐N4 sites in the oxygen reduction reaction (ORR) by electrochemical scanning tunneling microscopy (ECSTM). A well‐defined model catalytic system with explicit and uniform iodine‐coordinated Fe‐N4 sites was constructed facilely by the self‐assembly of iron(II) phthalocyanine (FePc) on an I‐modified Au(111) surface. The electrocatalytic activity of FePc for the ORR shows notable enhancement with axial iodine ligands. The modulation of the electronic structure of Fe sites to evoke a higher spin configuration by axial iodine was evidenced. The interaction strength between oxygen‐containing species and active centers becomes weaker due to the presence of iodine ligands, and the reaction is thermodynamically preferable. Furthermore, the reaction dynamics of FePc on I/Au(111) were explicitly determined via in situ ECSTM potential pulse experiments. In contrast, axial atomic iodine was found inefficacious for improving the activity of Co‐N4 sites, and electron rearrangement was found to be marginal, demonstrating that adequate interactions between axial ligands and metal sites for optimizing electronic structures and catalytic behaviors are prerequisites for the impactful role of axial ligands.

Sleep deprivation stimulates adaptive thermogenesis by activating AMPK pathway in mice.

Sleep deprivation (SD) can affect the adaptive thermogenesis in laboratory rodents, but the molecular mechanism and the crosstalk with other organs remain largely unknown. In order to investigate the effects and mechanisms of SD on thermoregulation and energy metabolism, here we measured the changes of body weight, body fat mass, body temperature, resting metabolic rate (RMR), and thermogenic gene expression in brown adipose tissue (BAT), white adipose tissue (WAT), skeleton muscle and liver in C57BL/6J mice during 7-day SD with rotating rod sleep deprivation device. Results showed that compared with the control group, the body weight and body fat mass of SD mice were decreased and RMR of SD mice increased. The gene expression of Ampk, Pgc1α and Ucp1 which related to thermogenesis in BAT and WAT were significantly increased, and the expression of Ampk, Serca1, Serca2 and Ucp3 which related to thermogenesis in skeletal muscle were significantly increased in SD mice. Taken together, these data demonstrated that 7-day SD enhanced the adaptive thermogenesis in mice by activating AMPK, including the upregulation of the AMPK - PGC1α - UCP1 pathway in BAT, and the AMPK - UCP3 and SLN - SERCA pathway in skeleton muscle. Our data provide the molecular evidence for SD-stimulated adaptive thermogenesis and energy metabolism in small mammals.

Complete Chloroplast Genome of Crassula aquatica: Comparative Genomic Analysis and Phylogenetic Relationships

Background/Objectives: Crassula aquatica (L.) Schonl. is a very small annual plant growing along riverbanks. Chloroplast (cp) genomes, crucial for photosynthesis, are highly conserved and play a key role in understanding plant evolution. In this study, we conducted cp genome analysis of C. aquatica, aiming to elucidate its phylogenetic position and structural variations. We analyzed and described the features of the complete cp genome of C. aquatica and conducted comparative analysis with the cp genomes of closely related taxa. Rsults: The cp genome was 144,503 bp in length and exhibited the typical quadripartite structure, consisting of a large single-copy region (LSC; 77,993 bp), a small single-copy region (SSC; 16,784 bp), and two inverted repeats (24,863 bp). The cp genome of C. aquatica comprised 113 unique genes, including 79 protein-coding genes (PCGs), 30 tRNAs, and 4 rRNA genes. Comparative genomic analysis of 13 other Crassula species and six outgroups demonstrated highly conserved gene content and order among Crassula species. However, notable differences were observed, including the complete loss of the rpoC1 intron in C. aquatica and several closely related species, which may serve as a synapomorphic trait supporting the monophyly of the subgenus Disporocarpa. We analyzed the nucleotide diversity among 14 Crassula cp genomes and identified five highly variable regions (pi > 0.08) in the IGS regions. Phylogenetic analysis based on 78 PCGs confirmed the monophyly of Crassula and its division into two subgenera: Crassula and Disporocarpa. Although the phylogenetic tree supported the subgeneric classification system, the sectional classification system requires reassessment. Conclusions: In this study, we conducted a comparative analysis of the cp genome of the genus Crassula. We inferred evolutionary trends within the Crassula cp genome and provided molecular evidence supporting the integration of the genus Tillaea into the genus Crassula. However, as this study does not represent all species within the genus Tillaea, further comprehensive phylogenetic analyses are requrired.

A novel Non-rodent animal model of hydrochloric acid-induced acute and chronic lung injury

Hydrochloric acid is one of the most prevalent and hazardous chemicals. Accidental spills occur in industrial plants or during transportation. Exposure to HCl can induce severe health impairment, including acute and chronic pulmonary diseases. We have previously described the molecular, structural, and functional aspects of the development of chronic lung injury and pulmonary fibrosis caused by intratracheal instillation of HCl in mice. Although mouse models of human disease have many advantages, rodents are evolutionary far from human and exhibit significant anatomical and physiological differences. Genetic and anatomic similarities between rabbits and humans are significantly higher. Rabbit models of HCl-induced lung injury have been used sparsely to evaluate acute lung injury. In this study, for the first time, we utilized rabbits as a model of HCl-induced pulmonary fibrosis and chronic lung injury. We present molecular, histological, and functional evidence that demonstrate the utility of using this model for studying new pharmaceutics against pulmonary fibrosis.

Molecular Evidence for Altered Angiogenesis in Neuroinflammation-Associated Schizophrenia and Bipolar Disorder Implicate an Abnormal Midbrain Blood-Brain Barrier.

Angiogenesis triggered by inflammation increases BBB permeability and facilitates macrophage transmigration. In the midbrain, we have discovered molecular alterations related to the blood-brain barrier (BBB), including endothelial cell changes associated with macrophage diapedesis, in neuroinflammatory schizophrenia and bipolar disorder, but changes in angiogenesis are yet to be reported. Hypothesis: We expected to discover molecular evidence of altered angiogenesis in the midbrain in individuals with schizophrenia and bipolar disorder compared to controls, with these changes more evident in "high" inflammation schizophrenia as compared to "low" inflammation. In a case-control post-mortem cohort including schizophrenia (n = 35), bipolar disorder (n = 35), and controls (n = 33), we measured mRNA (RT-PCR) and protein (multiplex immunoassays) and performed immunohistochemistry to determine levels and anatomical distribution of angiogenesis-related molecules in the ventral midbrain. We found large changes in angiogenesis factors in bipolar disorder high inflammatory subgroup (increased angiopoietin-2 and SERPINE1 mRNAs, but decreased angiopoietin-1, angiopoietin-2, andTEK receptor proteins). In schizophreniahigh inflammatory subgroup, we found a robust increase in SERPINE1 mRNA and protein levels. However, we found no significant changes in angiopoietins in schizophrenia. We found that VEGFA mRNA level was increased in high inflammation schizophrenia, but only reached statistical significance compared to one low inflammatory subgroup. Thus, angiogenesis signaling pathways appeared to be involved in the BBB alterations when inflammation is also present in the midbrain of schizophrenia and bipolar disorder, with increased levels of SERPINE1 in schizophrenia high inflammatory subgroup and with a putative suppression of angiopoietin signaling in bipolar disorder high inflammatory subgroup.

Lost in synonymy: Integrative species delimitation reveals two unrecognized species of Southern Asian tree squirrels (Rodentia: Sciuridae: Callosciurinae)

Abstract We present a comprehensive integrative taxonomic review of Callosciurus caniceps and Tamiops mcclellandii as they are currently defined. This review combines published molecular evidence, craniodental morphometrics, pelage and bacular variation, evaluations of potential hybrid zones using museum specimens and citizen science photographs, and, for C. caniceps, bioacoustic evidence. Our findings lead to the recognition of two species that had been lost in synonymy and highlight future perspectives on species delimitation in Sciuridae. By comparing phenotypic differentiation across climatic and vegetation transitions and contextualizing our results with the evolutionary history of our study systems, we provide insights into distribution, ecogeographical patterns, and speciation drivers in Southeast Asian vertebrates.

Photobiomodulation mitigates diastolic dysfunction, reduces perivascular fibrosis and inflammation, and enhances mitochondrial metabolism and angiogenesis in a mouse model of HFpEF

Abstract Introduction Emerging molecular evidence implicates mitochondrial dysfunction, systemic inflammation, and capillary rarefaction as pivotal contributors to the pathophysiology of HFpEF. Photobiomodulation (PBM) has garnered attention for its potential to augment mitochondrial metabolism, attenuate inflammation, and foster angiogenesis. This investigation aims to delineate the effects of PBM on the pathophysiology of HFpEF in a murine model. Methods To induce HFpEF, 3-month-old male C57BL/6 mice were subjected to a '2-hit model' involving a high-fat diet combined with the vasoconstrictor nitric oxide synthase inhibitor, N(G)-Nitro-L-arginine methyl ester (L-NAME). PBM therapy was administered transdermally over the cardiac region thrice weekly for a duration of 10 weeks using a 904nm super-pulsed laser, with each session lasting 90 seconds and delivering an energy density of 4.2J/cm^2 per diode to a cohort of 12 mice. A sham group of 13 mice underwent identical procedures with the laser deactivated. An additional control group of 10 mice was maintained on a standard diet without any interventions. Echocardiographic assessments were performed at the outset, 7th and 10th week time. Following the 10-week period, mice were euthanized, their hearts harvested and serum withdrawn for further analysis. Results Echocardiography of PBM-treated animals demonstrated significant attenuation of deterioration of diastolic parameters (E’) compared to the sham-treated control group (E’[mm/sec], baseline-vs-10w: PBM, -32.1±5.9-vs- -27.9±6.4; sham, -32.9±8.6-vs- -22.5±5.7; p=0.037). Histological analysis of the cardiac tissue involving picrosirius red staining for collagen revealed that the HFpEF model induced a significant increase in the standardized adventitial collagen compared to controls, which was attenuated by PBM (control 3.0±1.6; HFpEF sham: 3.9±1.8; PBM: 3.4±1.9, p=0.008 control vs sham, p=0.055 PBM vs control; p=0.396 control vs PBM). Multiplex analysis of HFpEF mice showed significantly higher levels of inflammatory chemokines than controls that were attenuated by PBM (mean±SD log[pg/ml] of the chemokine (C-X-C motif) ligand-1 (CXCL-1): PBM=2.52±0.29, Control=2.14±0.35; monocyte chemotactic protein (MCP-1): PBM=0.87±0.46, HFpEF=1.5±0.58, Control=0.8±0.6; p<0.001 and p=0.003 respectively). RNA sequencing revealed 551 genes with significant differential expression (DE) between PBM and Controls (adjusted p-value [padj]<0.1). Ingenuity Pathway Analysis (IPA) analysis revealed that PBM upregulated genes pertaining to mitochondrial metabolism and down-regulated genes active in mitochondrial dysfunction (qPCR [GAPDH] 2–∆∆Ct fold change PBM/Sham: Ndufv2=3.7, Atp5md=3.5, Dnhd1=0.5). Relevant DE genes were validated by quantitative real-time PCR (qPCR). Conclusions PBM attenuated Diastolic dysfunction, fibrosis, pro-inflammatory chemokine secretion and upregulates mitochondrial metabolism in a mouse model of HFpEF.Study designResults

Unraveling the molecular determinants of a rare human mitochondrial disorder caused by the P144L mutation of FDX2.

Episodic mitochondrial myopathy with or without optic atrophy and reversible leukoencephalopathy (MEOAL) is a rare, orphan autosomal recessive disorder caused by mutations in ferredoxin-2 (FDX2), which is a [2Fe-2S] cluster-binding protein participating in the formation of iron-sulfur clusters in mitochondria. In this biosynthetic pathway, FDX2 works as electron donor to promote the assembly of both [2Fe-2S] and [4Fe-4S] clusters. A recently identified missense mutation of MEOAL is the homozygous mutation c.431C>T (p.P144L) described in six patients from two unrelated families. This mutation alters a highly conserved proline residue located in a loop of FDX2 that is distant from the [2Fe-2S] cluster. How this Pro to Leu substitution damages iron-sulfur cluster biosynthesis is unknown. In this work, we have first compared the structural, dynamic, cluster binding and redox properties of WT and P144L [2Fe-2S] FDX2 to have clues on how the pathogenic P144L mutation can perturb the FDX2 function. Then, we have investigated the interaction of both WT and P144L [2Fe-2S] FDX2 with its physiological electron donor, ferredoxin reductase FDXR, comparing their electron transfer efficiency and protein-protein recognition patterns. Overall, the data indicate that the pathogenic P144L mutation negatively affects the FDXR-dependent electron transfer pathway from NADPH to FDX2, thereby reducing the capacity of FDX2 in assembling both [2Fe-2S] and [4Fe-4S] clusters. Our study also provided solid molecular evidences on the functional role of the C-terminal tail of FDX2 in the electron transfer between FDX2 and FDXR.

Multigene phylogenetics of Sargassum (Phaeophyceae) revealed low molecular diversity in contrast to high morphological variability in the NE Atlantic Ocean.

Sargassum species play a key role in habitat formation in tropical and subtropical regions; however, species identification has been hampered by the phenological plasticity exhibited in response to environmental conditions and life history. Molecular phylogenetics has challenged taxa circumscriptions and proven critical in delimiting species in this genus. Yet, the Atlantic species of Sargassum remain poorly understood, and recent studies have shown low molecular diversity between the species in the NW Atlantic. Here, we expand the taxon sampling to the NE Atlantic to assess the diversity of Sargassum in the Atlantic basin, based on a comprehensive morphological and multigene approach. We selected genes commonly used in delineating species in this genus (ITS2, rbcLS, cox3, mtsp) and explored additional markers (cox2, nad6, psbC, clpC, atpB) to infer the phylogenetic relationships between the morphospecies observed in the NE Atlantic. Phylogenetic analyses using single-gene and multigene alignments including 185 new sequences confirmed the low molecular diversity and supported the distinction of a single clade in Sargassum section Sargassum of N Atlantic benthic species. In contrast, morphological analysis resulted in the identification of 10 species and three new morphospecies that we described here but opt not to equate to species until further molecular evidence is available. Our results were congruent with previous findings from the NW Atlantic and highlight the morphological and ecological diversity of Sargassum in the Atlantic. These results suggest a recent colonization and incipient speciation of Sargassum in the Atlantic basin and showcase the need of further high-throughput analyses.

Experimental infection of ringtail possums (Pseudocheirus peregrinus) with Mycobacterium ulcerans, the agent of Buruli ulcer

Buruli ulcer (BU) is a necrotizing disease of skin and soft tissue caused by the bacterium Mycobacterium ulcerans (MU). In Australia, where the disease is emerging in new geographic areas and human case numbers are increasing, native possum species act as reservoir hosts. To better understand the life history of MU in one of its natural hosts, we conducted intra-dermal challenge of six wild caught, MU-naïve common ringtail possums (Pseudocheirus peregrinus). All six animals developed BU disease consistent with that observed in naturally infected ringtail possums. Time to ulceration varied between 49 and 77 days (mean = 61.8 days). Molecular evidence of systemic infection was detected in five animals and was supported by consistent histopathological findings in four animals. Pathological findings included random, multifocal, granulomatous hepatitis in four possums, one of which also had a mild, multifocal, interstitial granulomatous pneumonia. Acid-fast bacilli were only evident in inflammatory foci beyond the primary inoculation site in one possum. The ringtail possum model of MU infection is an important tool for the investigation of bacterial transmission dynamics, pathogenesis and immune response in a natural host. Data from this model may improve disease risk modelling and help identify intervention points to stop zoonotic transmission and disease spread.

Multiple introduction events expand the range of the invasive brown alga Rugulopteryx okamurae to northern Spain

The brown alga Rugulopteryx okamurae is one of the most recent and aggressive marine invaders known, with a non-native range that spans from the western Mediterranean to the southern Iberian Peninsula, and the European Macaronesian archipelagos. Here, we provide the first record of its presence at three disjunct areas from northern Spain, supported by morphological and molecular evidence. Rugulopteryx okamurae was discovered in the port of Bilbao in the northeastern Atlantic of Spain, more than 1200 km away from the nearest invaded location previously known. It was also found at two separate areas in northwestern Spain: San Amaro, a small inlet at the entrance of the port of A Coruña, and Monteferro-Playa América, a site close to the port of Vigo. In two areas, R. okamurae was found to cover up to 100 % of the bottom, suggesting that it may behave as an aggressive invader in northern Spain. Our findings confirm the propensity of R. okamurae for long-distance dispersal and reveal that this invader is expanding its range northwards along the European Atlantic coast.

A new agaricoid species of Porotheleum (Porotheleaceae, Agaricales) from Shandong Province, China

Porotheleum microsporum, a new agaricoid species of the genus Porotheleum (Porotheleaceae, Agaricales), was found growing on the live bark of Salix babylonica in Quancheng Park (Ji’nan City, Shangdong Province, China). This discovery, a testament to our rigorous research, is described and illustrated based on morphological, molecular, and ecological evidence. Our phylogenetic analyses based on DNA sequences of the internal transcribed spacers 1 and 2 with the 5.8S rDNA (ITS) and the large subunit of nuclear ribosomal DNA (LSU) support the inclusion of this new species in the genus Porotheleum as it forms an independent clade in the phylogenetic tree. Morphologically, P. microsporum differs from any known species by combining a pale-yellow pileus with an ochre-yellow center and relatively small basidiospores.

Salvia tainingensis, a neglected new species of Lamiaceae from Fujian Province, China

A species could not be known to the world of science if it not be discovered and described, considerable number of new species are neglected due to the plant itself and some other external restricted factors. On the basis of morphological and molecular evidences, a new species previously identified as Salvia scapiformis is now described and illustrated as Salvia tainingensis, named after Taining County in Fujian Province, where it was discovered. S. tainingensis does not closely resemble S. scapiformis, but is more similar to Salvia glabrifolia and Salvia daiguii, which were once identified as S. scapiformis, in terms of leaf shape, apex, base, corolla color, and connective length. However, it can be easily distinguished from the latter two by its thinly leathery leaves, serrate leaf margins, densely villous or glabrescent leaf abaxial, and the X-shaped arrangement of connective and filament.

Locoregional event or dyssynchronous distant metastasis: clinicopathological and molecular analysis of contralateral axillary lymph node metastasis in breast cancer patients.

Contralateral axillary lymph node metastasis (CAM) is a rare clinical condition in patients with breast cancer (BC). CAM can be either a locoregional event or a distant metastasis. Molecular application for clonal evolution in BC has not been reported in CAM cases. We studied six patients with CAM with clinical, pathological and/or molecular evidence of distant metastasis; those patients had poor outcomes. Two cases with molecular analysis of paired primary and CAM established clonal evolution of the CAM with its corresponding primary with additional molecular alteration, increased tumour mutation burden, and copy number variations (CNVs) in the CAMs. Four cases containing alterations from genes potentially modulate chromatin organization, supporting chromatin and subsequent transcriptional signature changes are essential in CAM. Molecular analysis is critical to establish the connection between CAM and its primary counterpart. Distant CAM shows clonal evolution compared with its corresponding primary with additional molecular alterations, increased mutation burden and/or copy number variations. CAM should be evaluated individually and handled in a personalized fashion. Evidence of a true metastatic CAM can be supported by distant metastasis to other organs, specific morphological features and/or clonal evolution.