Hemophagocytic lymphohistiocytosis and histiocytic necrotizing lymphadenitis secondary to hemodialysis catheter-related bloodstream infection caused by Corynebacterium Striatum.

BACKGROUND: We herein described the coexistence of hemophagocytic lymphohistiocytosis (HLH) and histiocytic necrotizing lymphadenitis, alternatively known as the Kikuchi disease (KD), secondary to hemodialysis catheter-related bloodstream infection (BSI) caused by Corynebacterium striatum. CASE PRESENTATION: A patient on maintenance hemodialysis had developed persistent fever and Corynebacterium striatum was subsequently identified from the culture of both catheter tip and peripheral blood. During mitigation of the BSI, however, his fever was unabated and ensuing workup further found thrombocytopenia, hyperferritinemia, hypertriglyceridemia, low NK cell activity and a surge in serum CD25 levels. Moreover, biopsy of the bone marrow and lymph node detected histopathological evidence of hemophagocytosis and KD, respectively. Upon these abnormalities, the title-bound diagnosis was considered and the patient was eventually recovered from the treatment of dexamethasone instead of antibiotics. Consistently, aberrations in his serum CD25 levels and NK cell activity had subsided two months after discharge. CONCLUSIONS: Arguably, this encounter offered a unique chance to unravel the principal pathogenic cascade in immunobiology that made the three entities one disease continuum. As such, our work may add new understandings of HLH and/or KD secondary to severe infections in general and excessive release of cytokines in particular among patients with kidney diseases. The resultant early diagnosis is crucial to initiate appropriate treatment and improve the survival of patients with these challenging and potentially life-threatening disorders.

Circ_0008529 Contributes to Renal Tubular Cell Dysfunction in High Glucose Stress via miR-185-5p/SMAD2 Pathway in Diabetic Nephropathy.

Circular RNA (circRNA) is key regulator of diabetic nephropathy (DN) progression. However, the role of circ_0008529 in DN progression remains to be better deciphered. Cell viability, cell cycle, apoptosis and inflammation were measured by MTS assay, flow cytometry and corresponding assay kits. RT-qPCR was used to assess the expression of circ_0008529, miR-185-5p and SMAD family member 2 (SMAD2). Also, western blotting was performed to measure protein expression. Target relationship was validated by RNA pull-down assay, dual-luciferase reporter assay and RNA immunoprecipitation assay. Urinary exosome was isolated using ultracentrifugation method and identified by transmission electron microscopy. Receiver operating characteristic curve was used to analyze the diagnostic value of circ_0008529 in DN patients. Circ_0008529 and SMAD2 were upregulated, while miR-185-5p was downregulated in high glucose (HG)-induced renal tubular HK-2 cells. Under HG treatment, cell viability and cell cycle process were suppressed, while apoptosis, inflammation and extracellular matrix accumulation were enhanced. However, interfering circ_0008529 could attenuate HG-induced effects, and this protection was abated by miR-185 inhibition or SMAD2 re-expression. Mechanically, circ_0008529 and SMAD2 were competing endogenous RNAs for miR-185-5p via target binding, and circ_0008529 regulated SMAD2 expression via miR-185-5p. Notably, circ_0008529 expression was upregulated in urinary exosomes of DN patients, and showed diagnostic value (Sensitivity: 70.21%; Specificity: 86.67%). Circ_0008529 might be a potential target for DN, which regulated DN progression via miR-185-5p/SMAD2 pathway.

Challenge of evolving Klebsiella pneumoniae infection in patients on hemodialysis: from the classic strain to the carbapenem-resistant hypervirulent one.

Loss of renal function may render hemodialysis patients more susceptible to infectious diseases, which is the second of all-causes mortality in this population. In addition to infection caused by the classic Klebsiella pneumoniae (cKp), however, hemodialysis staffs are now facing new challenge with growing prevalence of the carbapenem-resistant Kp (CR-Kp) and hypervirulent Kp (hvKp) as they are respectively associated with increased drug-resistance and virulence. We therefore chose to share our recent experience in treating severe infections either caused (cKp, CR-Kp, hvKp) or complicated (CR-hvKp) by these strains in hemodialysis patients. Based upon yet beyond published works, we further came up with the detection of intracranial lesion, novel diagnostic approach using unique biomarkers followed by selection of appropriate antibiotics, management of metastasic abscesses and bracing for the most lethal scenario in the order of cKp, CR-Kp, hvKp and CR-hvKp, respectively. Since reports of complicated hvKp infection in hemodialysis patients were rare, we also discussed in details this clinical entity focusing on its epidemiology, mechanism of increased virulence and involvement of the arteriovenous fistula as insidious source of persistent septicemia. By covering the full spectrum of clinically relevant Kp stains specifically from the viewpoint of nephrology, our work had highlighted the importance of infection control in uremic state and vice versa. As such, it may greatly raise the awareness of dialysis staffs against the challenge of evolving Klebsiella pneumoniae infection in hemodialysis patients and expeditiously reach a higher degree of readiness which was proved to be the key determinant of ultimate survival.

First Report of Bacterial Leaf Spot on Tobacco Caused by Pseudomonas psychrotolerans in China.

Tobacco (Nicotiana tabacum) is an economically important crop, and its productivity is challenged due to pathogen infection. In 2020 and 2021, a previously uncharacterized disease was observed on field grown tobacco (Variety NC102) in Zhucheng City, Shandong Province, China (119°7'14" E, 36°0'58" N), where tobacco has been grown for decades. The disease can be found throughout the growth period of tobacco and mainly occurred from fast growing period (about 13-16 leaves) to leaf maturity stage. In severely diseased areas, the incidence rate can reach 100%. The symptoms first began as chlorotic water stain like small spots, then the spots merged into larger irregular necrotic maculae around the chlorotic halos. Small pieces of symptomatic leaves from 10 different infected plants were collected for pathogen isolation. The small pieces of discolored leaves were surface sterilized with 75% ethanol for 40s and washed with sterile water for three times. The sterilized leaves were ground with a glass rod with 1mL sterile water, and 100 µL suspensions were spread on nutrient agar medium then incubated at 28oC for 48 hours. Yellow round colonies with undulating edges were showed up on nutrient agar medium 48 hours later. Three isolates were randomly picked up from each of the 10 plates for subsequent analysis. After purification and culture on nutrient agar plate, the 16S rRNA gene of the 30 isolates were amplified with primers 27F and 1492R and the amplicons were sequenced and analyzed by sequence alignment. The sequence alignment results showed that the 16S rRNA nucleotide identity of the 30 isolates were 100%. One typical isolate named ZC5 was selected for subsequent analysis, and the resulting 16S rRNA sequence was deposited at GenBank, NCBI under accession OK092624. The 16S rRNA sequence identity with those of P. psychrotolerans strain K3-2 (KY882083) and M3-1 (KY882120) were 100%, respectively. The phenotypic analysis by Biolog Gen Ⅲ indicated that the bacterial isolate (ZC5) showed highest similarity (98.3%) with strain Pseudomonas oryzihabitans. P. oryzihabitans and P. psychrotolerans have a high degree of homology in the phylogenetic relationship based on the phylogenetic analysis of three concatenated sequences of gyrB, rpoB and rpoD genes (Mulet et al. 2010). The gyrB (ON462356), rpoB (ON462355), rpoD (ON462357) gene of isolate ZC5were also amplified and sequenced by using primers gyrB-For/gyrB-Rev, rpoB-For/rpoB-Rev and rpoD-For/rpoD-Rev (Hauser et al. 2004), respectively. While P. psychrotolerans and P. oryzihabitans form the same clade in phylogenies, strains of P. psychrotolerans do form a unique sub-clade. Isolate ZC5 clustered more closely with the type strain of P. psychrotolerans LMG 21977 in the phylogenetic tree. Therefore, based on the concatenated sequences of three genes (gyrB, rpoB and rpoD), the isolate ZC5 was confirmed as P. psychrotolerans. Based on morphological, Biolog characteristics and phylogenetic analysis, the isolate was identified as P. psychrotolerans. The tobacco plants at fast growing stage were selected for pathogenicity tests. Pathogenicity tests were conducted by injecting 10 µL bacterial suspension (108cfu/mL) of ZC5 into tobacco leaves with a syringe. Sterile water was inoculated into the tobacco leaves in the same way as the control. Six plants were selected for pathogenicity tests each time and five leaves of each tobacco plant were inoculated, and the tests were repeated three times. To simulate disease conditions in the natural environment, the inoculated plants were moved outdoors. The average temperature was 32°C during the day and 20°C at night. To maintain humidity, the tobacco leaves were sprayed with water every two days. Symptoms appeared on the pathogen inoculated leaves seven days after inoculation, whereas the control treatment remained symptomless. The pathogens were reisolated from diseased leaves and identified as P. psychrotolerans based on morphological, molecular and phylogenetic analysis, which fulfilled Koch's postulates. To our knowledge, this is the first report of tobacco bacterial leaf spot caused by P. psychrotolerans.

Suppression of lncRNA GAS6-AS2 alleviates sepsis-related acute kidney injury through regulating the miR-136-5p/OXSR1 axis in vitro and in vivo.

Acute kidney injury (AKI) is a common complication of sepsis and increase morbidity and mortality. Long non-coding RNA (LncRNA) GAS6-AS2 was related to inflammation and apoptosis in different diseases by regulating miRNAs and downstream genes, but its role in AKI remains unclear. Thus, we speculated that GAS6-AS2 might function in sepsis-related AKI via regulating target genes. Here, LPS or CLP was used to establish in vitro or in vivo sepsis-related AKI model. The interactions between GAS6-AS2 and miR-136-5p, and miR-136-5p and OXSR1, were validated by luciferase reporter assay, RNA pull-down, or RIP assay. Cell apoptosis was determined by flow cytometry, Western blotting, or IHC. The kidney injury was evaluated by H&E staining. The expression of GAS6-AS2, miR-136-5p, and OXSR1 was determined by qRT-PCR or Western blotting. We found that GAS6-AS2 was up-regulated in LPS-treated HK2 cells and the CLP-induced rat model. In vitro, GAS6-AS2 knockdown decreased cleaved caspase-3 and bax expression and increased bcl-2 expression. The levels of TNF-α, IL-1ß, and IL-6 were reduced by GAS6-AS2 down-regulation. GAS6-AS2 knockdown ameliorated oxidative stress in the cells, as indicated by the reduced ROS and MDA levels and the elevated SOD level. In vivo, GAS6-AS2 down-regulation decreased urinary NGAL and Kim-1 levels and serum sCr and BUN levels, and H&E proved that the kidney injury was alleviated. GAS6-AS2 knockdown also reduced apoptosis, inflammation, and oxidation induced by CLP in vivo. Mechanically, GAS6-AS2 sponged miR-136-5p which targeted OXSR1. Overall, lncRNA GAS6-AS2 knockdown has the potential to ameliorate sepsis-related AKI, and the mechanism is related to miR-136-5p/OXSR1 axis.

Sequencing and phylogenetic characterization of a novel RNA virus in Arma chinensis.

The complete genome of a novel virus from Arma chinensis was determined by RNA sequencing and rapid amplification of cDNA ends. This virus has a single-stranded RNA genome of 10,540 nucleotides (nt) excluding the poly(A) tail. Two non-overlapping open reading frames (ORFs) in the sense direction were predicted: one long ORF at the 5' end of the genome (6,219 nt) that encodes a polypeptide of 2,072 amino acids (aa), and one short ORF at the 3' end of the genome (3,033 nt) that encodes a polypeptide of 1,010 aa. Phylogenetic analysis indicated that the virus clusters within a large cluster of currently unidentified picorna-like viruses with a high bootstrap value. We named the virus isolate Arma chinensis picorna-like virus 1 (AcPV-1). The prevalence of AcPV-1 infection in samples of Arma chinensis from the wild was at a low level (5.48%, 8 positives in 146 samples). Keywords: Arma chinensis; genomic characterization; phylogenetic analysis; Arma chinensis picorna-like virus 1; prevalence.

Analogous and Diverse Functions of APSES-Type Transcription Factors in the Morphogenesis of the Entomopathogenic Fungus Metarhizium rileyi.

APSES-type transcription factors (TFs) have analogous and diverse functions in the regulation of fungal morphogenesis processes. However, little is known about these functions in microsclerotium formation. In this study, we characterized two orthologous APSES genes (MrStuA and MrXbp) in the entomopathogenic fungus Metarhizium rileyi Deletion of either MrStuA or MrXbp impaired dimorphic transition, conidiation, fungal virulence, and microsclerotium formation. Compared with the wild-type strain, ΔMrStuA and ΔMrXbp mutants were hypersensitive to thermal and oxidative stress. Furthermore, transcriptome sequencing analysis revealed that MrStuA and MrXbp independently regulate their own distinctive subsets of signaling pathways during dimorphic transition and microsclerotium formation, but they also show an overlapping regulation of genes during these two distinct morphogenesis processes. These results provide a global insight into vital roles of MrStuA and MrXbp in M. rileyi and aid in dissection of the interacting regulatory mechanisms of dimorphism transition and microsclerotium development.IMPORTANCE Transcription factors (TFs) are core components of the signaling pathway and play an important role in transcriptional regulation of gene expression during fungal morphogenesis processes. A prevailing theory suggests an interplay between different TFs regulating microsclerotial differentiation; however, the persisting issue remains that these interplay mechanisms are not clear. Here, we analyzed two members of the APSES-type TFs in Metarhizium rileyi using a gene deletion strategy and transcriptome analysis. Mutants were significantly impaired in microsclerotium formation and dimorphic transition. Transcriptome analysis provided evidence for interacting regulatory mechanisms by the two TFs in microsclerotium formation and dimorphic transition. Furthermore, we investigated their overlapping roles in mediating the expression of genes required for different fungal morphogenesis processes. Characterization of TFs in this study will aid in dissecting the interplay between regulatory mechanisms in fungal morphogenesis processes.

Sequencing and phylogenetic characterization of a novel Polerovirus from Nicotiana tabacum.

In this study, we reported the complete genome of a novel Polerovirus, named Tobacco yellow virus (TYV), which can be transmitted by Myzus persicae. TYV had a single-stranded RNA genome of 5735 nucleotides in length and contained six putative open reading frames (ORFs). Phylogenetic analysis with whole genome nucleotide sequences and amino acid sequences deduced from the conserved domain of the RNA-dependent RNA polymerase, clustered TYV with Potato leafroll virus from the genus Polerovirus with high bootstrap values. However, TYV clustered with Brassica yellow virus using amino acid sequences deduced from the conserved domain of the coat protein. Taken together with the identities between ORFs in TYV and related ORFs in species from Polerovirus, our results strongly suggested TYV is a novel species of the genus Polerovirus.

Sequencing and phylogenetic characterization of a novel RNA virus genome from Harmonia axyridis.

As a natural predator of many insect pests on its native Asian range, Harmonia axyridis remains amongst the insects whose pathogenic or beneficial microorganisms are yet to be studied. The genome nucleotide (nt) and amino acid sequences of open reading frames (ORFs) of the novel RNA virus were identified. Neighbor-joining (NJ) were constructed using MEGA7 software packages with nt sequences and conserved amino acid sequences of predicted RNA-dependent RNA polymerase (RdRp).The complete genome of a novel virus named Harmonia axyridis virus 1 was determined by RNA-seq and rapid amplification of cDNA ends from H. axyridis, which had a single-stranded RNA genome of 8868 nts in length and contains two putative ORFs. ORF1 encodes a polypeptide of 2182 amino acids, which contained conserved domains for 2 picornavirus-like capsid proteins and one RNA helicase. ORF2 encodes a polypeptide of 655 amino acids, which contained 1 RdRp domain. Phylogenetic analysis of whole genome nt sequences and RdRp deduced amino acid sequences suggested that the virus clustered with several unclassified Hubei picorna-like virus. To our knowledge, this is the first full annotated genome of a novel member of the unclassified group of RNA viruses, infecting H. axyridis in natural field conditions.

Biocontrol of tobacco black shank disease (Phytophthora nicotianae) by Bacillus velezensis Ba168.

Tobacco black shank (TBS) caused by Phytophthora nicotianae is destructive to almost all tobacco cultivars and is widespread in many tobacco-growing countries. Through lab study and field test, we isolated plant growth-promoting rhizobacteria (PGPR) strain Ba168 which is a promising biocontrol strain of TBS. Ba168 was isolated from 168 soil samples and identified as Bacillus velezensis by its genetic and phenotypic characteristics. A susceptibility test indicated that the P. nicotianae antagonistic materials of Ba168 in extracellular metabolites were composed of effective and stable proteins/peptides. P. nicotianae's growth was suppressed by the ammonium sulfate precipitation of Ba168 culture filtrates (ASPBa) at a minimum inhibitory concentration of 5 µg/mL. Extracellular conductivity, pH, and the wet/dry weights of P. nicotianae's mycelia, along with scanning electron microscope analysis, suggested that Ba168-derived proteins/peptides could effectively inhibit P. nicotianae by causing irreversible damage to its cell walls and membranes. Protein identification of ASPBa supported these results and identified many key proteins responsible for various biocontrol-related pathways. Field assays of TBS control efficacy of many PGPRs and agrochemicals showed that all PGPR preparations reduced the disease index of tobacco, but Ba168 was the most effective. These results demonstrated the importance of Bacillus-derived proteins/peptides in the inhibition of P. nicotianae through irreversible damage to its cell wall and membrane; and the effectiveness of PGPR strain B. velezensis Ba168 for biocontrol of the soil-borne disease caused by P. nicotianae.

First report of Colletotrichum chlorophyti causing peanut anthracnose in China.

Peanut (Arachis hypogaea L.) is an important oil crop - mainly in Shandong and Henan Provinces, in China. In 2018-2019, the occurrence of a black spot disease on the leaves and stems were found in two fields (a total of 10 ha) in Niulingguanzhuang village of Yishui county in Linyi, Shandong Province, China, and 20% to 40% of plants were infected, thereby reducing the amount of marketable product. Natural symptoms were circular, dark brown-to-black lesions (2-6 mm in diameter) and coalescent necrosis on leaves and black necrosis in stems. Six symptomatic leaves and stems collected from six plants of two fields, were used for isolation. Portions of infected tissue were surfaced-sterilized with 0.5% NaClO for two minutes, 70% alcohol for 30 seconds and washed twice with sterile water. The tissues were placed on potato dextrose agar (PDA) at 28℃ for 5 days. Felt or fleece forming colonies about 4 cm in diameter that were circular, flat, and with dark center and white narrow margins were formed. Three isolates (LSJF-4, HSGF, HSJF) were purified and one culture (LSJF-4) was deposited in the China General Microbiological Culture Collection Center (CGMCC, NO. 3.19617). The chlamydospore were dark brown, verruculose, organized in chains and clusters, yellow-brown to black-brown color, and mostly spherical, 10.3 - 18.4 µm × 8.3-11.4 µm (n = 50, av. 12.6 ± 2.1× 10.1 ± 1.1 µm). The conidia were colorless, crescent or sickle-shaped, with one acute end and one blunt end. They were 16.7 - 24.3 µm × 3.6 - 5.5 µm (n= 50, av. 21.2 ± 1.7× 4.5 ± 0.36 µm), with dark brown, straight, septate setae, 40.3 - 86.9 µm × 3.6-5.5 µm (n=50, av. 70.0 ± 19.1 × 4.5 ± 0.7 µm) in size. Setae were straight, dark brown. These morphological characteristics of our isolates were identical to Colletotrichum chlorophyti (Damm et al. 2009). The internal transcribed spacer (ITS, ITS1/ITS4) rDNA of three fungus (LSJF-4, HSGF, HSJF) were amplified using PCR and sequenced, as described by Damm et al (2009). The sequencing results (GenBank Accession No. MK796409, MN756650, MN756651) were submitted to the GenBankand blast analysis showed that it had 99.0% identity with those of Colletotrichum chlorophyti (No. GU227894). Actin (ACT, ACT-512F/ACT-783R), beta-tubulin (Tub2, T1/T2 ), Glyceraldehyde-3-phosphate dehydrogenase (GAPDH, GDF1/GDR1), chitin synthase (CHS1, CHS-79f/CHS-345R), histone (H3, Cy1H3F/ Cy1H3R) of LSJF-4 (Nos. MN688800, MN688797, MN097811, MN688799 and MN688798, respectively) (Damm et al. 2009) were also sequenced and BLAST. The results showed high identity of all the five sequences to the CGMCC 3.19617 (Nos. GU228286/GU228287 in GAPDH = 99.6%, GU228384/GU22385 in CHS1 = 96.8%, GU228090/GU228091 in H3 = 99.0%, GU227992/GU227993 in ACT = 98.7%, and GU228188/GU228189 in Tub2 = 98.8%). Therefore, isolate CGMCC 3.19617 was identified as C. chlorophyti based on morphological and molecular characteristics. To determine pathogenicity, a conidial suspension containing 106 conidia/mL was sprayed on leaves and stems of six 40-day old peanut seedlings. Three control plants were similarly sprayed with sterile water. All treated plants were kept moist (>85% relative humidity) at 30℃ for 48 h in darkness, and then kept in 60% relative humidity and 28℃ conditions. Two weeks post-inoculation, small, dark, near elliptical lesions appeared on inoculated leaves and stems, which were similar to those naturally infected plants. While controls remained symptomless. C. chlorophyti was reisolated from infected tissues and identified based on the previous methods, fulfilling Koch's postulates. The test was repeated twice. C. chlorophyti has been reported as a pathogen associated with soybean anthracnose isolated from the Netherlands (Damm et al. 2009). C. chlorophyti was first detected on soybean plants imported from Uruguay (Li et al. 2017) in China. To our knowledge, this is the first report of C. chlorophyti causing peanut anthracnose in China. C. chlorophyti may be a new threat to Leguminous plant species, especially peanuts plants. We proposed paying close attention to taking necessary control meatures.

Preference of the aphid Myzus persicae (Hemiptera: Aphididae) for tobacco plants at specific stages of potato virus Y infection.

Potato virus Y (PVY) is a common pathogen affecting agricultural production worldwide and is mainly transmitted by Myzus persicae in a non-persistent manner. Insect-borne plant viruses can modify the abundance, performance, and behavior of their vectors by altering host plant features; however, most studies have overlooked the fact that the dynamic progression of virus infection in plants can have variable effects on their vectors. We addressed this point in the present study by dividing the PVY infection process in tobacco into three stages (early state, steady state and late state); delineated by viral copy number. We then compared the differential effects of PVY-infected tobacco (Nicotiana tabacum) plants on the host selection and feeding behavior of M. persicae. We used Y-shaped olfactory apparatus and electrical penetration graph (EPG) methods to evaluate host selection and feeding behavior, respectively. Interestingly, we found that PVY-infected plants at the steady state attracted more aphids than healthy plants, whereas no differences were observed for those at the early and late states. In terms of feeding behavior, intracellular punctures (closely related to PVY acquisition and transmission) were more abundant on PVY-infected tobacco plants at the early and steady states of infection than in uninfected plants. These results indicate that PVY-infected host plants can alter the host selection and feeding behavior of aphids in a stage-dependent manner, which is an important consideration when studying the interactions among host plants, viruses, and insect vectors.

The bZIP transcriptional factor activator protein-1 regulates Metarhizium rileyi morphology and mediates microsclerotia formation.

Internal oxidative stress can trigger microsclerotia (MS) formation of Metarhizium rileyi in liquid culture. Activator protein 1 (AP1) is a transcription factor and an important determinant of the response to oxidative stress. To investigate how M. rileyi responds to internal oxidative stress and how MS development is regulated, the Mrap1 gene was characterized. Mrap1 was highly expressed during periods of invasive hyphal growth and in response to changing culture conditions during MS development. Compared with the wild-type and complemented strains, ΔMrap1 mutants exhibited various defects in aerial hyphal growth, yeast-to-hypha transition, and conidia and MS formation. ΔMrap1 mutants also displayed sensitivity to oxidative stress, were morphologically abnormal, and responded differently to oxidative stress during MS development. ΔMrap1 mutants had significantly reduced conidial (74-82%) and MS (99%) yields. Insect bioassays revealed that ΔMrap1 mutants displayed reduced virulence in topical (43-76%) and injection (45-70%) bioassays. Moreover, the ability of ΔMrap1 mutants to grow out of the cuticle was reduced due to impaired conidiation on the host cadaver. Digital gene expression profiling revealed that genes involved in antioxidation, pigment biosynthesis, and ion transport were regulated by Mrap1 during MS development. Taken together, our results confirm the importance of Mrap1 in vegetative growth, conidia and MS formation, and virulence.

Prenylated Diphenyl Ethers from the Marine Algal-Derived Endophytic Fungus Aspergillus tennesseensis.

Considerable attention has been paid to marine derived endophytic fungi, owing to their capacity to produce novel secondary metabolites with potent bioactivities. In this study, two new compounds with a prenylated diphenyl ether structure-diorcinol L (1) and (R)-diorcinol B (2)-were isolated from the marine algal-derived endophytic fungus Aspergillus tennesseensis, along with seven known compounds: (S)-diorcinol B (3), 9-acetyldiorcinol B (4), diorcinol C (5), diorcinol D (6), diorcinol E (7), diorcinol J (8), and a dihydrobenzofuran derivative 9. Their structures were elucidated by extensive NMR spectroscopy studies. Compound 2 represents the first example of an R-configuration in the prenylated moiety. All these isolated compounds were examined for antimicrobial and cytotoxic activities. Compounds 1⁻9 exhibited antimicrobial activities against some human- and plant-pathogenic microbes with MIC values ranging from 2 to 64 µg/mL. Moreover, compound 9 displayed considerable inhibitory activity against the THP-1 cell line in vitro, with an IC50 value of 7.0 µg/mL.

A novel single-stranded RNA virus in Nesidiocoris tenuis.

The complete genome sequence of a novel single-stranded RNA virus in Nesidiocoris tenuis was determined by RNA-seq and rapid amplification of cDNA ends (RACE) methodologies and was named N. tenuis virus 1. The genomic RNA was 3970 nucleotides (nt) in length and contained two putative open reading frames (ORFs). ORF1 encoded a polypeptide with 283 amino acids containing a viral (superfamily 1) RNA helicase (Hel) domain, and ORF2 encoded a polypeptide with 294 amino acids containing an RNA-dependent RNA polymerase (RdRP) domain. Phylogenetic analysis using the deduced amino acid sequences indicated that the N. tenuis virus 1 clustered with Blackford virus; however, the low bootstrap values and unique genomic structure suggested that the virus is a prototype of a new type of unclassified viruses. The prevalence of N. tenuis virus 1 infection in field populations of N. tenuis differed between three locations, with 28.32% of the 113 sampled individuals testing positive for the virus.

Characterization and Distribution Analysis of a Densovirus Infecting Myzus persicae nicotianae (Hemiptera: Aphididae).

Densoviruses (DVs) are a group of viruses that contain a linear single-stranded DNA genome between 4­6 kb in length. Herein, we report a DV with a 5,480-nt genome, isolated from tobacco aphid (Myzus persicae nicotianae Blackman), named MpnDV. Unlike the genome of M. persicae densovirus (MpDV), which possesses five open reading frames (ORFs), the genome of MpnDV contains four putative ORFs­the nonstructural protein 1 (NS1) and NS2 from MpnDV are 98- and 52-amino acids longer than those of MpDV, respectively, at the N-terminus, and the capsid proteins (VP) are 102 amino acids longer at the C-terminus than those of MpDV. Mapping of the MpnDV transcripts by RACE method indicated that the ORF of NS2 started at nt 340 and the right two putative ORFs were combined together by deleting two introns, one of 95 bp located at nt 2,932­3,026 and the other of 145 bp located at nt 4,715­4,859, suggesting transcript mapping was necessary for analyzing of genome organization. Alignment analysis indicated that MpnDV shows 97% sequence identity with MpDV, and that the shortened ORFs resulted from nucleotide indels, suggesting MpnDV and MpDV were two isolates of the same virus. Thus, MpnDV and MpDV clustered together in a tree-based analysis. The prevalence of MpnDV infection in wild populations of tobacco aphids differed among 29 locations; 34% of the 622 individuals sampled were positive. The genome organization, transcript strategy, and widespread distribution in wild populations suggest that MpnDV might possess a biological function different from that of MpDV.

An AIL/IL-based liquid/liquid extraction system for the purification of His-tagged proteins.

A sorbent based on affinity ionic liquid (AIL), triazacyclononane-ionic liquid, was synthesized, characterized, and applied to the extraction of histidine (His)-tagged proteins from aqueous buffer to ionic liquid (IL) phase. The adsorbed His-tagged proteins could be back-extracted from the IL phase to the aqueous buffer with an imidazole solution. The specific binding of His-tagged proteins with AIL/IL could be affected by a few factors including the ionic strength and coordinated metal ions. In the case of His-tagged enhanced green fluorescent protein (EGFP), the maximum binding capacity of Cu(2+)-AIL/IL reached 2.58 µg/µmol under the optimized adsorption conditions. The eluted His-tagged EGFP kept fluorescent and remained active through the purification process. Moreover, a tandem extraction process successively using Cu(2+)-AIL/IL and Zn(2+)-AIL/IL systems was developed, which was proven very efficient to obtain the ultimate protein with a purity of about 90 %. An effective reclamation method for the AIL/IL extraction system was further established. The sorbent could be easily regenerated by removing metal ions with EDTA and the followed reimmobilization of metal ions. Easy handling of the presented M(2+)-AIL/IL system and highly specific ability to absorb His-tagged proteins make it attractive and potentially applicable in biomolecular separation.

Pulmonary tumor embolism in a maintenance hemodialysis patient with hepatocellular carcinoma.

Patients with chronic kidney disease are already at an increased risk for pulmonary embolism, since loss of renal function rendered a procoagulant state. Further, malignant tumor is a well-established risk factor for pulmonary thromboembolism. Alternatively, occlusion of the pulmonary vasculature by tumor cells per se and associated thrombi may mimic thromboembolic disease. By comparison, however, report of pulmonary tumor embolism (PTE) in patients on maintenance hemodialysis (MHD) is exceedingly rare. A less vigilant clinician may have otherwise treated this situation as fluid overload or thromboembolic disorder. We herein described in an MHD patient such an unusual case of PTE, which was diagnosed by contrast-enhanced CT and PET/CT. As such, our work may expand the knowledge reserve of dialysis staffs about this rare complication of malignancy.

Knowledge, Attitudes, and Practice of General Practitioners Toward Screening of Age-Related Hearing Loss in Community: A Cross-Sectional Study in Shanghai, China.

Background: Age-related hearing loss (ARHL) is experiencing a continuously rising in prevalence among the elderly worldwide. General practitioners (GPs) may have a unique position in its community detection and management. Objective: This study aims to assess the KAP of GPs regarding ARHL through questionnaire, to investigate the role of them in the management and to propose strategies for the hearing screening within the community. Methods: An online survey was administered to 1173 GPs, selected from 56 community health centers (CHCs) in Shanghai during April to June 2022. A scale endorsed by a panel of multidisciplinary experts was used to assess knowledge (7 items), attitudes (12 items), and practice (10 items). A mean score was computed and converted into a scale ranging from 0 to 100. Odds ratios (ORs) were calculated for potential predictors of higher levels of KAP scores (with mean value as a cutoff point) through logistic modelling. Results: A total of 1022 GPs completed the questionnaire with response rate 87.13%. The average scores are 69.90 ± 32.27, 66.09 ± 7.15, and 59.89 ± 21.99 for Knowledge, attitude, and practice, respectively. 24.3% of participants achieve a complete score of knowledge, whereas 5.48% receive zero. 11.6% consider ARHL as not a disease. Above 30.0% are not familiar with the screening tool. 10.8% refuse to undergo hearing screening. Higher levels of compliance in practice are found in the participants with higher levels of knowledge (OR=1.409, p=0.000) and more favorable attitude (OR=1.028, p=0.000). Male (OR=0.708, p=0.036) is associated with lower levels of attitudes. Conclusion: GPs have a low level of ARHL knowledge, a lack of positive attitude towards the detection and management of it, and lower awareness in practice. Further research is required to gain a more comprehensive understanding of the attitudes held by GPs and explore more accessibility strategies.

Pulmonary cryptococcosis closely mimicking lung cancer in a membranous nephropathy patient taking calcineurin inhibitor.

In patients with membranous nephropathy (MN), malignancy may be either the underlying disease or results of immunosuppressive therapy which may also lead to opportunistic infections including the pulmonary cryptococcosis. On CT scan, nodule is the most common feature in pulmonary cryptococcosis and it can mimic lung cancer both clinically and radiologically. Therefore, pulmonary nodular lesions caused by cryptococcosis may be easily misdiagnosed and require unnecessary surgical treatment. As such, we herein presented an isolated subpleural solitary nodule with satellite lesion that closely mimicked lung cancer on both contrast-enhanced computed tomography (CT) scan and 18F-fluorodeoxyglucose positron emission tomography (FDG-PET)/CT in an MN patient under long-term tacrolimus regimen. Cryptococcosis was ascertained by the finding of oval thick-walled yeast on histopathology of the lung biopsy specimen taken during the Argon-Helium cryotherapy. Further, the pulmonary lesions progressively dissipated after antifungal treatment. Arguably, our experience may help clinicians in general and nephrologists in particular with a better understanding of the cryptococcal infection manifesting as pulmonary nodule(s) in the MN patients and contribute to more efficacious differential diagnosis against the lung cancer.