Acutalibacter caecimuris sp. nov., Acutalibacter intestini sp. nov. and Neglectibacter caecimuris sp. nov., three novel species of the family Oscillospiraceae isolated from caecal contents of C57BL/6J mice.

The intestines of mice are colonized by diverse, as-yet-uncultivated bacteria. In this report, we describe the isolation, culture, genotypic and phenotypic characterization, as well as taxonomic classification of three novel anaerobic bacterial strains derived from the caecal contents of C57BL/6J male mice. According to the phenotypic and genotype-based polyphasic taxonomy, we propose three novel species within the family Oscillospiraceae. They are Acutalibacter caecimuris sp. nov. (type strain M00118T=CGMCC 1.18042T=KCTC 25739T), Acutalibacter intestini sp. nov. (type strain M00204T=CGMCC 1.18044T=KCTC 25741T) and Neglectibacter caecimuris sp. nov. (type strain M00184T=CGMCC 1.18043T=KCTC 25740T).

Lacticaseibacillus rhamnosus Strains for Alleviation of Irritable Bowel Disease and Chronic Fatigue Syndrome.

Lacticaseibacillus rhamnosus is applied as a probiotic to alleviate various metabolic, gastrointestinal, and psychological symptoms and diseases, and its probiotic effectiveness is strain-specific. In this study, we obtained 21 strains of Ls. rhamnosus, and their genomes were sequenced. We defined the pan- and core-genomes of Ls. rhamnosus. Phenotypes such as the assimilation of carbohydrates and antibiotic resistance were experimentally characterized and associated with genome annotations. Nine strains were selected and tested for growth rates, tolerance to acidity/alkalinity and bile acids, the production of short-chain fatty acids, and competition with pathogenic microbes. Strains WL11 and WL17 were targeted as potential probiotics and were applied in mouse model tests for the alleviation of chronic fatigue syndrome (CFS) and irritable bowel syndrome (IBS). The results showed that WL11 and WL17 effectively alleviated slow body weight gain, anxiety, poor memory, and cognitive impairment in CFS mouse models. They also reduced the expression of pro-inflammatory factors, such as TNF-α and IL-6, and alleviated intestinal peristalsis, visceral hypersensitivity, and anxiety-like behavior in IBS mouse models. This study reports new Ls. rhamnosus strain resources and their effect on alleviation of both IBS and CFS symptoms with mouse models; the probiotic functions of those strains in human patients remain to be further tested.

Ecoclimate drivers shape virome diversity in a globally invasive tick species.

Spillovers of viruses from animals to humans occur more frequently under warmer conditions, particularly arboviruses. The invasive tick species Haemaphysalis longicornis, the Asian longhorned tick, poses a significant public health threat due to its global expansion and its potential to carry a wide range of pathogens. We analyzed meta-transcriptomic data from 3595 adult H. longicornis ticks collected between 2016 and 2019 in 22 provinces across China encompassing diverse ecological conditions. Generalized additive modeling revealed that climate factors exerted a stronger influence on the virome of H. longicornis than other ecological factors, such as ecotypes, distance to coastline, animal host, tick gender, and antiviral immunity. To understand how climate changes drive the tick virome, we performed a mechanistic investigation using causality inference with emphasis on the significance of this process for public health. Our findings demonstrated that higher temperatures and lower relative humidity/precipitation contribute to variations in animal host diversity, leading to increased diversity of the tick virome, particularly the evenness of vertebrate-associated viruses. These findings may explain the evolution of tick-borne viruses into generalists across multiple hosts, thereby increasing the probability of spillover events involving tick-borne pathogens. Deep learning projections have indicated that the diversity of the H. longicornis virome is expected to increase in 81.9% of regions under the SSP8.5 scenario from 2019 to 2030. Extension of surveillance should be implemented to avert the spread of tick-borne diseases.

Molecular phylogeny of the family Rhabdiasidae (Nematoda: Rhabditida), with morphology, genetic characterization and mitochondrial genomes of Rhabdias kafunata and R. bufonis.

BACKGROUND: The family Rhabdiasidae (Nematoda: Rhabditida) is a globally distributed group of nematode parasites, with over 110 species parasitic mainly in amphibians and reptiles. However, the systematic position of the family Rhabdiasidae in the order Rhabditida remains unsolved, and the evolutionary relationships among its genera are still unclear. Moreover, the present knowledge of the mitochondrial genomes of rhabdiasids remains limited. METHODS: Two rhabdiasid species: Rhabdias kafunata Sata, Takeuchi & Nakano, 2020 and R. bufonis (Schrank, 1788) collected from the Asiatic toad Bufo gargarizans Cantor (Amphibia: Anura) in China, were identified based on morphology (light and scanning electron microscopy) and molecular characterization (sequencing of the nuclear 28S and ITS regions and mitochondrial cox1 and 12S genes). The complete mitochondrial genomes of R. kafunata and R. bufonis were also sequenced and annotated for the first time. Moreover, phylogenetic analyses based on the amino acid sequences of 12 protein-coding genes (PCGs) of the mitochondrial genomes were performed to clarify the systematic position of the family Rhabdiasidae in the order Rhabditida using maximum likelihood (ML) and Bayesian inference (BI). The phylogenetic analyses based on the 28S + ITS sequences, were also inferred to assess the evolutionary relationships among the genera within Rhabdiasidae. RESULTS: The detailed morphology of the cephalic structures, vulva and eggs in R. kafunata and R. bufonis was revealed using scanning electron microscopy (SEM) for the first time. The characterization of 28S and ITS regions of R. kafunata was reported for the first time. The mitogenomes of R. kafunata and R. bufonis are 15,437 bp and 15,128 bp long, respectively, and both contain 36 genes, including 12 PCGs (missing atp8). Comparative mitogenomics revealed that the gene arrangement of R. kafunata and R. bufonis is different from all of the currently available mitogenomes of nematodes. Phylogenetic analyses based on the ITS + 28S data showed Neoentomelas and Kurilonema as sister lineages, and supported the monophyly of Entomelas, Pneumonema, Serpentirhabdias and Rhabdias. Mitochondrial phylogenomic results supported Rhabdiasidae as a member of the superfamily Rhabditoidea in the suborder Rhabditina, and its occurrance as sister to the family Rhabditidae. CONCLUSIONS: The complete mitochondrial genome of R. kafunata and R. bufonis were reported for the first time, and two new gene arrangements of mitogenomes in Nematoda were revealed. Mitogenomic phylogenetic results indicated that the family Rhabdiasidae is a member of Rhabditoidea in Rhabditina, and is closely related to Rhabditidae. Molecular phylogenies based on the ITS + 28S sequence data supported the validity of Kurilonema, and showed that Kurilonema is sister to Neoentomelas. The present phylogenetic results also indicated that the ancestors of rhabdiasids seem to have initially infected reptiles, then spreading to amphibians.

A Phase-Field Study of Spinodal Decomposition Impeded by Irradiation in U-Mo and U-Mo-Zr Alloys.

The phase-field method, coupled with the micro-elastic model and irradiation-induced cascade mixing model, has been employed to investigate the spinodal decomposition in U-Mo and U-Mo-Zr alloys. The microstructure evolution of U-Mo or U-Mo-Zr alloys under different initial conditions, such as the alloy composition, aging temperature and irradiation intensity, were simulated to study the effect of cascade mixing on the miscibility gap, morphology and volume fraction of the decomposed phases. The simulation results demonstrate that irradiation-induced cascade mixing impedes the process of spinodal decomposition, and that irradiation shrinks the composition range of the miscibility gap in the alloys. Irradiation-induced cascade mixing slows down the anisotropic growth rate of the spinodal decomposition, yet this phenomenon can be weakened with increasing aging temperature. Adding an appropriate amount of Zr to a U-Mo alloy can effectively prevent the contraction of the miscibility gap caused by irradiation.

Extensive genetic diversity of severe fever with thrombocytopenia syndrome virus circulating in Hubei Province, China, 2018-2022.

Severe fever with thrombocytopenia syndrome virus (SFTSV), an etiological agent causing febrile human disease was identified as an emerging tick-borne bunyavirus. The clinical disease characteristics and case fatality rates of SFTSV may vary across distinct regions and among different variant genotypes. From 2018 to 2022, we surveyed and recruited 202 severe fever with thrombocytopenia syndrome (SFTS) patients in Hubei Province, a high-incidence area of the epidemic, and conducted timely and systematic research on the disease characteristics, SFTSV diversity, and the correlation between virus genome variation and clinical diseases. Our study identified at least 6 genotypes of SFTSV prevalent in Hubei Province based on the analysis of the S, M, and L genome sequences of 88 virus strains. Strikingly, the dominant genotype of SFTSV was found to change during the years, indicating a dynamic shift in viral genetic diversity in the region. Phylogenetic analysis revealed the genetic exchange of Hubei SFTSV strains was relatively frequent, including 3 reassortment strains and 8 recombination strains. Despite the limited sample size, SFTSV C1 genotype may be associated with higher mortality compared to the other four genotypes, and the serum amyloid A (SAA) level, an inflammatory biomarker, was significantly elevated in these patients. Overall, our data summarize the disease characteristics of SFTSV in Hubei Province, highlight the profound changes in viral genetic diversity, and indicate the need for in-depth monitoring and exploration of the relationship between viral mutations and disease severity.

In vivo self-assembled small RNA targets H19 lncRNA for the treatment of colorectal cancer.

The majority of molecularly targeted therapies in clinical use target disease-related proteins, but only a small fraction (∼1.5%) of human genome is protein-coding region. Considering that ∼70% of human genome is transcribed to noncoding RNAs, targeting noncoding RNAs rather than protein-coding RNAs can significantly expand the proportion of human genome that can be manipulated. H19 long noncoding RNA (lncRNA) is aberrantly expressed in a variety of cancer types and actively contributes to multiple steps of tumorigenesis. Therefore, we selected H19 as a representative target and designed synthetic anti-H19 construct for the self-assembly and delivery of anti-H19 small RNA (sRNA) to prevent colorectal cancer development and metastasis based on the natural ability of the host liver to package sRNA-encapsulating small extracellular vesicles (sEVs) and the endogenous circulating sEVs to transfer sRNA. As anticipated, the synthetic anti-H19 construct successfully generated anti-H19 sRNA-encapsulating sEVs and exhibited high silencing efficiency on H19 lncRNA in an ex vivo model. In orthotopic and lung metastasis mouse models of colorectal cancer, the anti-H19 construct exhibited significantly superior therapeutic efficacy over 5-fluorouracil (5-Fu) in preventing primary tumor growth and lung metastasis. Particularly, the anti-H19 sRNA-encapsulating sEVs were generated in a nontoxic, nonimmunogenic and biocompatible manner. In summary, this study demonstrates that the in vivo self-assembled anti-H19 sRNA can serve as a new therapeutic agent for colorectal cancer.

Effect of different aging treatments on the transport of nano-biochar in saturated porous media.

Widely used for soil amendment, carbon sequestration, and remediation of contaminated soils, biochars (BCs) inevitably produce a large number of nanoparticles with relatively high mobility. Geochemical aging alters chemical structure of these nanoparticles and thus affect their colloidal aggregation and transport behavior. In this study, the transport of ramie derived nano-BCs (after ball-milling) was investigated by different aging treatments (i.e., photo (PBC) and chemical aging (NBC)) as well as the managing BC under different physicochemical factors (i.e., flow rates, ionic strengths (IS), pH, and coexisting cations). Consequences of the column experiments indicated aging promoted the mobility of the nano-BCs. Compared to the nonaging BC, consequences of spectroscopic analysis demonstrated the aging BCs exhibited a number of tiny corrosion pores. Both of these aging treatments contribute to a more negative zeta potential and a higher dispersion stability of the nano-BCs, which is caused by the abundance of O-functional groups. Also the specific surface area and mesoporous volume of both aging BCs increased significantly, with the increase being more pronounced for NBC. The breakthrough curves (BTCs) obtained for the three nano-BCs were modelled by the advection-dispersion equation (ADE), which included first-order deposition and release terms. The ADE revealed high mobility of aging BCs, which meant their retention in saturated porous media was reduced. This work contributes to a comprehensive understanding of the transport of aging nano-BCs in the environment.

New insights into the impact of microbiome on horizontal and vertical transmission of a tick-borne pathogen.

BACKGROUND: The impact of host skin microbiome on horizontal transmission of tick-borne pathogens , and of pathogen associated transstadial and transovarial changes in tick microbiome are largely unknown, but are important to control increasingly emerging tick-borne diseases worldwide. METHODS: Focusing on a rickettsiosis pathogen, Rickettsia raoultii, we used R. raoultii-positive and R. raoultii-negative Dermacentor spp. tick colonies to study the involvement of skin microbiota in cutaneous infection with rickettsiae in laboratory mice, and the function of the tick microbiome on maintenance of rickettsiae through all tick developmental stages (eggs, larvae, nymphs, adults) over two generations. RESULTS: We observed changes in the skin bacteria community, such as Chlamydia, not only associated with rickettsial colonization but also with tick feeding on skin. The diversity of skin microbiome differed between paired tick-bitten and un-bitten sites. For vertical transmission, significant differences in the tick microbiota between pathogenic rickettsia-positive and -negative tick chorts was observed across all developmental stages at least over two generations, which appeared to be a common pattern not only for R. raoultii but also for another pathogenic species, Candidatus Rickettsia tarasevichiae. More importantly, bacterial differences were complemented by functional shifts primed for genetic information processing during blood feeding. Specifically, the differences in tick microbiome gene repertoire between pathogenic Rickettsia-positive and -negative progenies were enriched in pathways associated with metabolism and hormone signals during vertical transmission. CONCLUSIONS: We demonstrate that host skin microbiome might be a new factor determining the transmission of rickettsial pathogens through ticks. While pathogenic rickettsiae infect vertebrate hosts during blood-feeding by the tick, they may also manipulate the maturation of the tick through changing the functional potential of its microbiota over the tick's life stages. The findings here might spur the development of new-generation control methods for ticks and tick-borne pathogens. Video Abstract.

Skin infectome of patients with a tick bite history.

Introduction: Ticks are the most important obligate blood-feeding vectors of human pathogens. With the advance of high-throughput sequencing, more and more bacterial community and virome in tick has been reported, which seems to pose a great threat to people. Methods: A total of 14 skin specimens collected from tick-bite patients with mild to severe symptoms were analyzed through meta-transcriptomic sequencings. Results: Four bacteria genera were both detected in the skins and ticks, including Pseudomonas, Acinetobacter, Corynebacterium and Propionibacterium, and three tick-associated viruses, Jingmen tick virus (JMTV), Bole tick virus 4 (BLTV4) and Deer tick mononegavirales-like virus (DTMV) were identified in the skin samples. Except of known pathogens such as pathogenic rickettsia, Coxiella burnetii and JMTV, we suggest Roseomonas cervicalis and BLTV4 as potential new agents amplified in the skins and then disseminated into the blood. As early as 1 day after a tick-bite, these pathogens can transmit to skins and at most four ones can co-infect in skins. Discussion: Advances in sequencing technologies have revealed that the diversity of tick microbiome and virome goes far beyond our previous understanding. This report not only identifies three new potential pathogens in humans but also shows that the skin barrier is vital in preventing horizontal transmissions of tick-associated bacteria or virus communities to the host. It is the first research on patients' skin infectome after a tick bite and demonstrates that more attention should be paid to the cutaneous response to prevent tick-borne illness.

Causal relationship between serum uric acid and abnormal blood pressure based on the panel model study: A 5-year cohort study.

BACKGROUND AND AIMS: To investigate the relationship between elevated serum uric acid (SUA) levels and blood pressure (BP). METHODS AND RESULTS: Based on the Beijing Health Management Cohort, 5276 health examination people were enrolled. Cross-lagged model was used to explore the relationship between SUA levels and blood pressure. The results showed: (1) increased SUA and increased systolic blood pressure (SBP): ① The path coefficients from baseline SUA to follow-up SBP were statistically significant in both the general population (ß = 0.034, P < 0.05) and men (ß = 0.048, P < 0.05). The path coefficients from baseline SBP to follow-up SUA were not statistically significant in either the general population (ß = 0.010, P > 0.05) or men (ß = 0.011, P > 0.05). ② The path coefficients from baseline SUA to follow-up SBP and from baseline SBP to follow-up SUA were not statistically significant in women with BMI ≥ 25 kg/m2 and BMI < 25 kg/m2. (2) Increased SUA and diastolic blood pressure (DBP): ① There was no statistical significance between the path coefficients from baseline DBP to follow-up SUA and the path coefficients from baseline SUA to follow-up DBP. ② In men and women, BMI ≥ 25 kg/m2 and BMI < 25 kg/m2, the path coefficients from baseline DBP to follow-up SUA and from baseline SUA to follow-up DBP were not statistically significant. CONCLUSIONS: SUA can increase blood pressure in the general male population; no reverse time sequence relationship was found. The temporal relationships between SUA levels and SBP abnormalities were different in the sex and BMI subgroups. No bidirectional causal temporal relationship was found between SUA elevation and DBP abnormality.

Metavirome of 31 tick species provides a compendium of 1,801 RNA virus genomes.

The increasing prevalence and expanding distribution of tick-borne viruses globally have raised health concerns, but the full repertoire of the tick virome has not been assessed. We sequenced the meta-transcriptomes of 31 different tick species in the Ixodidae and Argasidae families from across mainland China, and identified 724 RNA viruses with distinctive virome compositions among genera. A total of 1,801 assembled and complete or nearly complete viral genomes revealed an extensive diversity of genome architectures of tick-associated viruses, highlighting ticks as a reservoir of RNA viruses. We examined the phylogenies of different virus families to investigate virome evolution and found that the most diverse tick-associated viruses are positive-strand RNA virus families that demonstrate more ancient divergence than other arboviruses. Tick-specific viruses are often associated with only a few tick species, whereas virus clades that can infect vertebrates are found in a wider range of tick species. We hypothesize that tick viruses can exhibit both 'specialist' and 'generalist' evolutionary trends. We hope that our virome dataset will enable much-needed research on vertebrate-pathogenic tick-associated viruses.

Austraplectana extranes sp. n. (Nematoda: Cosmocercoidea) from Australian amphibians Mixophyes carbinensis Mahony, Donnellan, Richards & McDonald (Anura: Myobatrachidae) and Rhinella marina (Linnaeus) (Anura: Bufonidae).

The genus Austraplectana Baker, 1981 is a poorly known group of cosmocercoid nematodes. In the present study, a new species of Austraplectana, A. extranes sp. n., was described using both light and scanning electron microscopy, based on specimens collected from the carbine barred frog Mixophyes carbinensis Mahony, Donnellan, Richards & McDonald (Anura: Myobatrachidae) and the cane toad Rhinella marina (Linnaeus) (Anura: Bufonidae) in Australia. Austraplectana extranes sp. n. can be easily distinguished from its congener by the much longer and different morphology of tails in both sexes, the different number and arrangement of caudal papillae, the presence of single precloacal median heart-shaped papilla and large posterior protuberance with cuticular comb-like fringe in male.

Atmospheric HONO formation during and after the Spring Festival holidays in a coastal city of China.

Nitrous acid (HONO) is an important source of hydrogen oxides (HOx), which affects air quality, the atmospheric oxidation capacity, and human health. Here, we present ambient measurements of the HONO concentrations in Zhuhai, a coastal city in Southern China, from February 7 to March 15, 2021. The campaign was classified into two periods during (P1) and after (P2) the Spring Festival holidays. The average HONO mixing ratio during P2 (1.19 ± 0.85 ppbv) was much higher than that during P1 (0.24 ± 0.18 ppbv), likely due to the contribution of homogeneous HONO formation. During nighttime, the heterogeneous conversion rate during P2 (0.0089/hr) was considerably higher than that during P1 (0.0057/hr), suggesting a higher heterogeneous NO2 conversion potential. However, the heterogeneous NO2 conversion was the dominant way during P1 with a high percentage of 88%, while comparable ratios of heterogeneous and homogeneous formation were found (54% vs. 46%) during P2, indicating that the homogeneous formation was also important during P2. During daytime, homogeneous reaction was the major known pathway, with a contribution of 16% during P1 and 27% during P2, leaving large unknown HONO sources which reasonably correlated with the photo-enhanced NO2 conversion. Two case scenarios were additionally explored, showing that there might be a primary emission source during one scenario (February 17-18) and vehicle emissions might be the major unknown HONO source for another scenario (March 3-5). The results suggest that large unknown daytime sources still exist which need more future ambient and laboratory studies.

Cruise observation of the marine atmosphere and ship emissions in South China Sea: Aerosol composition, sources, and the aging process.

Marine atmospheric aerosols impact the global climate and biogeochemical cycles. However, how the composition, sources, and aging of these aerosols affect the above processes has not been thoroughly studied. Here, we conducted ship-based measurements in the northern South China Sea to investigate the chemical composition and aging of aerosols from various sources during the summer of 2019. Separate measurements were conducted at the bow (marine environment) and stern (cooking, smoking, and engine exhaust) of the ship. Source apportionment of organic aerosols (OAs) was conducted using positive matrix factorization (PMF) and trajectory models. The results showed that ship exhaust and coastal submicron particles were composed of comparable sulfate and organic fractions (both approximately 43%), distinct from the sulfate-dominated particles in the marine atmosphere (52-77%). PMF using the multilinear engine-2 solver identified five factors for the stern sampling period: hydrocarbon-like OA (HOA-I, 9%), slightly oxidized HOA (HOA-II, 25%), cooking OA (COA, 13%), cigarette smoke OA (CSOA, 4%), and low-volatility oxygenated OA (LV-OOA, 49%). The primary OAs (HOA-I/II + COA + CSOA), derived mostly from direct ship-related emissions, contributed to approximately half of the OAs, whereas the contribution from the highly aged marine atmosphere was only 20%. Notably, certain living-related emissions (i.e., COA and CSOA), which were often neglected in previous studies, might represent a considerable contribution to OA emissions from the ship. Four factors were identified for the bow sampling periods: HOA (13%), biomass burning OA (BBOA, 9%), semi-volatile OOA (7%), and LV-OOA (71%). The BBOAs from the Indo-China and Malay peninsulas were aged, converted to secondary organic aerosols (SOAs) during transport, and influenced by the combined photo-oxidation and liquid-phase reactions, indicating a substantial impact of BB on SOA formation. Our study highlights the influence of ship and inland emissions and their aging during transport on marine atmospheric aerosols.

Native species Maxvachonia chabaudi Mawson, 1972 (Nematoda: Cosmocercoidea) found in the invasive marine toad Rhinella marina (Linnaeus) (Anura: Bufonidae) in Australia.

The genus Maxvachonia Chabaud et Brygoo, 1960 (Ascaridomorpha: Cosmocercidae) is a poorly known group of parasitic nematodes. Species of Maxvachonia are native to Madagascar-Australo-Papuan Region, where they are known to parasitise frogs, snakes and skinks. Unfortunately, most of Maxvachonia species have been inadequately described. In the present study, we report the native species Maxvachonia chabaudi Mawson, 1972 from the intestine of the invasive marine toad Rhinella marina (Linnaeus) in Australia for the first time. We speculate that the marine toads infected with M. chabaudi are likely related to their eating skinks or the similarity in diet/habitat/ecology between the toad and the skinks. The detailed morphology of M. chabaudi was studied using light microscopy and, for the first time, scanning electron microscopy, based on the newly collected specimens. Some characters important for the specific diagnosis of M. chabaudi are reported for the first time, including each lip with distinct inner flanges, the location of vulva varying from anterior to posterior of the oesophageal bulb and the presence of single medio-ventral precloacal papilla. An identification key to the species of Maxvachonia is provided.

A Review of the Role of Ultrasound Radiomics and Its Application and Limitations in the Investigation of Thyroid Disease.

The incidence of thyroid disease has gradually increased in recent years. Conventional ultrasound is one of the most critical thyroid imaging methods, but it still has certain limitations. The use of B-model ultrasound (BMUS) diagnosis of thyroid disease will be affected by a doctors' clinical experience. The ultrasound radiomics is based on ultrasound images to delineate the region of interest (ROI), and then extract features to quantify the disease information contained in the image, which helps to analyze the correlation between the image and the clinical pathology of the disease. By building a powerful model, it can be used to diagnose benign and malignant thyroid nodules, predict lymph node status in thyroid cancer, analyze molecular biological characteristics, and predict the survival of thyroid cancer patients. At present, the application of ultrasound radiomics in the thyroid is pervasive. These ultrasound radiomics studies have further promoted the progress of ultrasonic technology in the field of thyroid disease. Clinicians should be familiar with the workflow of ultrasound radiomics and understand the application of this technology to the thyroid. In this article, we first describe the workflow of ultrasound radiomics, followed by an overview of the application of ultrasound radiomics to the thyroid. Finally, some current limitations of the technology and areas for future improvement are discussed. This article aims to review the role of ultrasound radiomics and its application and limitations in the investigation of thyroid disease.

Ginsenoside Rb1 inhibits astrocyte activation and promotes transfer of astrocytic mitochondria to neurons against ischemic stroke.

Astrocytes activation in response to stroke results in altered mitochondrial exchange with neurons. Ginsenoside Rb1is a major ginsenoside of Panax ginseng particularly known for its neuroprotective potential. This work aimed to investigate if Rb1 could rescue neurons from ischemic insult via astrocyte inactivation and mitochondrial transfer. We prepared conditioned astrocytes-derived medium for co-culture with neurons and examined the role of Rb1 in mitochondrial transfer from astrocytes to neurons. The neuroprotective potential of Rb1 was further confirmed in vivo using a mouse model of brain ischemia. In response to oxygen-glucose deprivation and reperfusion (OGD/R), astrocytes were reactivated and produced reactive oxygen species (ROS), an action that was blocked by Rb1. Mechanistically, Rb1 inhibited NADH dehydrogenase in mitochondrial complex I to block reverse electron transport-derived ROS production from complex I, and thus inactivated astrocytes to protect the mitochondria. Mitochondrial signal, mitochondrial membrane potential and ATP production detected in conditioned astrocyte-derived medium indicated that Rb1 protected functional mitochondria and facilitated their transfer. When neurons were injured by OGD/R insult, co-culturing with conditioned medium increased mitochondrial membrane potential and oxygen consumption rate within the neurons, indicating the protection conferred on them by Rb1 via mitochondrial transfer from astrocytes. Using the ischemic mouse brain model, CD38 knockdown in the cerebral ventricles diminished the neuroprotective effects of Rb1, providing evidence in support of the role of astrocyte mitochondrial transfer. Transient inhibition of mitochondrial complex I by Rb1 reduced mitochondrial ROS production and consequently avoided astrocyte activation. Astrocyte mitochondrial transfer therefore seemed a means by which Rb1 could promote neuronal survival and function. Different from the neurocentric view, these findings suggest the astrocytes may be a promising target for pharmacological interventions in ischemic brain injury.

Morphology, genetic characterization and molecular phylogeny of the poorly known nematode parasite Cissophyllus leytensis Tubangui & Villaamil, 1933 (Nematoda: Ascaridida) from the Philippine sailfin lizard Hydrosaurus pustulatus (Eschscholtz, 1829) (Reptilia: Squamata).

BACKGROUND: The genus Cissophyllus (Cosmocercoidea: Kathlaniidae) is a rare group of nematodes parasitic in turtles and lizards. To date, only four species have been reported in Asia and North America. However, most of them are inadequately described. The species Cissophyllus leytensis has never been reported since it was originally described by Tubangui and Villaamil in 1933 from the Philippine sailfin lizard Hydrosaurus pustulatus (Eschscholtz, 1829) (Reptilia: Squamata). Furthermore, the systematic status of Cissophyllus/Cissophyllinae in the family Kathlaniidae of the superfamily Cosmocercoidea remains under debate. METHODS: The detailed morphology of C. leytensis was studied using light microscopy (LM) and, for the first time, scanning electron microscopy (SEM), based on newly collected specimens from the type host H. pustulatus. Six different genetic markers, including nuclear sequences [small ribosomal subunit (18S), internal transcribed spacer (ITS) and large ribosomal subunit (28S)], plus mitochondrial genes [cytochrome c oxidase subunit 1 (cox1), cytochrome c oxidase subunit 2 (cox2) and 12S small subunit ribosomal RNA gene] of C. leytensis were sequenced. Additionally, in order to test the validity of the subfamily Cissophyllinae and clarify the phylogenetic relationships of Cissophyllus and the other genera in the family Kathlaniidae, phylogenetic analyses based on 18S + 28S and ITS sequence data were performed using maximum likelihood (ML) and Bayesian inference (BI) analyses, respectively. RESULTS: Our observations using LM and SEM revealed some previously unreported morphological features, necessitating the redescription of this poorly known species. The presence of remarkable morphological variation in the isthmus and the position of excretory pore among different individuals was found. Molecular analysis showed no intraspecific nucleotide divergence detected in the 18S, ITS, 28S, cox2 and 12S regions among different individuals of C. leytensis, but a low level of intraspecific genetic variation was found in the cox1 (0.52%). Our phylogenetic results showed the representatives of the Cosmocercoidea divided into four large clades (Cosmocerca + Aplectana + Cosmocercoides representing the family Cosmocercidae, Cruzia representing the subfamily Cruzinae of Kathlaniidae, Falcaustra + Cissophyllus + Megalobatrachonema representing the subfamily Kathlaniinae of Kathlaniidae, and Orientatractis + Rondonia representing the family Atractidae). The genus Cissophyllus clustered together with the genus Megalobatrachonema in both the ML and BI trees using ITS sequence data, but displayed a sister relationship to the genus Falcaustra in the ML tree and to the genera Falcaustra + Megalobatrachonema in the BI tree using 18S + 28S sequence data. CONCLUSIONS: Molecular phylogenetic results further confirmed that the family Kathlaniidae is not a monophyletic group. The subfamily Cruziinae should be moved from the hitherto-defined family Kathlaniidae and elevated as a separate family Cruziidae. The present phylogenetic results also negated the validity of the subfamily Cissophyllinae and supported the genus Cissophyllus assigned in the subfamily Kathlaniinae. Molecular analysis indicated that the morphological variation in the isthmus and position of excretory pore among different individuals should be considered as intraspecific variation. Moreover, some characters important for the specific diagnosis of C. leytensis are reported for the first time: the number of acuminate denticles (lamellae) on each lip, the chitinized pharynx with three flabellate pharyngeal plates, the presence of single medioventral precloacal papilla and the detailed morphology of caudal papillae. The present study is only the second record of C. leytensis.

The regulatory gene wblA is a target of the orphan response regulator OrrA in Streptomyces coelicolor.

Our previous study using transposon mutagenesis indicated that disruption of the putative response regulator gene orrA impacted antibiotic production in Streptomyces coelicolor. In this study, the role of OrrA was further characterized by comparing the phenotypes and transcriptomic profiles of the wild-type S. coelicolor strain M145 and ΔorrA, a strain with an inactivated orrA gene. Chromatin immunoprecipitation using a strain expressing OrrA fused with FLAG showed that OrrA binds the promoter of wblA, whose expression was downregulated in ΔorrA. The interaction of OrrA with the wblA promoter was further validated by a pull-down assay. Similar to ΔorrA, the deletion mutant of wblA (ΔwblA) was defective in development, and developmental genes were expressed at similar levels in ΔorrA and ΔwblA. Although both OrrA and WblA downregulated actinorhodin and undecylprodigiosin, their roles in regulation of the calcium-dependent antibiotic and yellow-pigmented type I polyketide differed. sco1375, a gene of unknown function, was identified as another OrrA target, and overexpression of either sco1375 or wblA in ΔorrA partially restored the wild-type phenotype, indicating that these genes mediate some of the effects of OrrA. This study revealed targets of OrrA and provided more insights into the role of the orphan response regulator OrrA in Streptomyces.