Identification and fungicide sensitivity of Microdochiumchrysopogonis (Ascomycota, Amphisphaeriaceae), a new species causing tar spot of Chrysopogonzizanioides in southern China.

Vetiver grass (Chrysopogonzizanioides) has received extensive attention in recent years due to its diverse applications in soil and water conservation, heavy metal remediation, as well as essential oil and phenolic acids extraction. In 2019, the emergence of tar spot disease on C.zizanioides was documented in Zhanjiang, Guangdong Province, China. Initially, the disease manifested as black ascomata embedded within leaf tissue, either scattered or clustered on leaf surfaces. Subsequently, these ascomata became surrounded by fisheye lesions, characterised by brown, elliptical, necrotic haloes, which eventually coalesced, resulting in leaf withering. Koch's postulates demonstrated that the fungus isolated from these lesions was the causal agent. Microscopic examination showed that the pathogen morphologically belonged to Microdochium. The phylogenetic tree inferred from the combined ITS, LSU, tub2 and rpb2 sequences revealed the three isolates including GDMCC 3.683, LNU-196 and LNU-197 to be a novel species of Microdochium. Combining the results of phylogenetic, pathogenicity and morphological analyses, we propose a new species named M.chrysopogonis as the causal agent of C.zizanioides in southern China. The optimum growth temperature for M.chrysopogonis was determined to be 30 °C. The in vitro fungicide sensitivity of M.chrysopogonis was determined using a mycelial growth assay. Four demethylation-inhibiting (DMI) fungicides, including difenoconazole, flusilazole, propiconazole and tebuconazole and one methyl benzimidazole carbamate (MBC) fungicide, carbendazim, were effective against M.chrysopogonis, with mean 50% effective concentration (EC50) values of 0.077, 0.011, 0.004, 0.024 and 0.007 µg/ml, respectively. These findings provide essential references for the precise diagnosis and effective management of M.chrysopogonis.

First report of postharvest fruit rot on passion fruit (Passiflora edulis) caused by Lasiodiplodia theobromae in Mainland China.

As an economically important tropical and subtropical fruit crop, passion fruit (Passiflora edulis Sims) is widely planted in many provinces of southern China. In April 2017, postharvest fruit rot was observed on 15% to 25% of passion fruit in several fruit markets of Zhanjiang City in Guangdong Province. Initial disease symptoms on infected fruit were irregular, brown, water-soaked lesions, which enlarged into large black and sunken patches. Lesions were usually covered with an abundance of little black dots (pycnidia) and black-gray hyphae. For the pathogen isolation, fifteen symptomatic fruit were randomly collected from three local markets. Fourteen single-spore fungal isolates with similar morphology ware isolated from the infected tissues. Two isolates (ZW 49-1 and ZW 50-1) were randomly selected to further study. The colonies on PDA were initially greyish-white and became dark-gray with age. Abundant globular and irregular pycnidia were observed after incubation at 25 °C for 3 weeks. The conidia of the fungus were initially hyaline, unicellular, apex rounded, thick-walled, and ellipsoid, becoming dark brown, bicellular with longitudinal striations at maturity, 26.4 ± 2.5 × 13.4 ± 1.2 µm (n = 50). The morphology of the fungus resembled Lasiodiplodia theobromae (Pat.) Griff. & Maubl. (Phillips et al. 2013). To confirm species identification, the partial internal transcribed spacer (ITS) region of rDNA, translation elongation factor-alpha (EF1-α) and ß-tubulin (TUB) gene were amplified from genomic DNA of the two isolates with the ITS1/ITS4, EF1-688F/EF1-986R, and Bt2a/Bt2b primers, respectively (Glass and Donaldson 1995; Alves et al. 2008; White et al. 1990). Base on the BLASTn analysis, the ITS (MT644473, MT644474), EF1-α (MT649210, MT649211) and TUB (MT649212, MT649213) sequences of both isolates were 100%, 99% and 100% similarity to the L. theobromae CBS 164.96 ex-type sequences in the NCBI database (AY640255, AY640258, and KU887532, respectively) (Phillips et al. 2013). For pathogenicity test, asymptomatic passion fruit were previously disinfested in 0.5 % sodium hypochlorite and superficially wounded with a sterile needle. Five-mm-diameter plugs with mycelial taken from 5-day-old PDA colonies were placed on the wounds. Sterile PDA plugs were used as negative controls. Each treatment had five replicates and the test was repeated twice. Fruit were maintained in plastic boxes to keep at 25°C for one week. One week after inoculation, gray mycelia had covered a majority of the fruit surface and caused a black, sunken rot. The inoculated fungus was reisolated and confirmed as L. theobromae by morphological characteristics. The mock inoculated fruit remained asymptomatic. The occurrence of fruit rot on passion fruit caused by L. theobromae was reported in Taiwan, China recently (Huang et al., 2019). To our knowledge, this is the first report of L. theobromae causing postharvest fruit rot on passion fruit in the Chinese mainland.

Identification of Subtype-Specific Metastasis-Related Genetic Signatures in Sarcoma.

Background: Sarcomas are heterogeneous rare malignancies constituting approximately 1% of all solid cancers in adults and including more than 70 histological and molecular subtypes with different pathological and clinical development characteristics. Method: We identified prognostic biomarkers of sarcomas by integrating clinical information and RNA-seq data from TCGA and GEO databases. In addition, results obtained from cell cycle, cell migration, and invasion assays were used to assess the capacity for Tanespimycin to inhibit the proliferation and metastasis of sarcoma. Results: Sarcoma samples (N = 536) were divided into four pathological subtypes including DL (dedifferentiated liposarcoma), LMS (leiomyosarcoma), UPS (undifferentiated pleomorphic sarcomas), and MFS (myxofibrosarcoma). RNA-seq expression profile data from the TCGA dataset were used to analyze differentially expressed genes (DEGs) within metastatic and non-metastatic samples of these four sarcoma pathological subtypes with DEGs defined as metastatic-related signatures (MRS). Prognostic analysis of MRS identified a group of genes significantly associated with prognosis in three pathological subtypes: DL, LMS, and UPS. ISG15, NUP50, PTTG1, SERPINE1, and TSR1 were found to be more likely associated with adverse prognosis. We also identified Tanespimycin as a drug exerting inhibitory effects on metastatic LMS subtype and therefore can serve a potential treatment for this type of sarcoma. Conclusions: These results provide new insights into the pathogenesis, diagnosis, treatment, and prognosis of sarcomas and provide new directions for further study of sarcoma.

DnaJA4 is involved in responses to hyperthermia by regulating the expression of F-actin in HaCaT cells.

BACKGROUND: Hyperthermia in combination with DnaJA4-knockout (KO) obviously affects the anti-viral immunity of HaCaT cells. The mechanisms of this process are not yet fully explored. However, it is known that DnaJA4 interacts with actin cytoskeleton after hyperthermia. Our aim was to investigate the effects of DnaJA4 on F-actin in HaCaT cells following hyperthermia. METHODS: Wild-type (WT) and DnaJA4-KO HaCaT cells were isolated at either 37°C (unheated) or 44°C (hyperthermia) for 30 min followed by testing under conditions of 37°C and assessing at 6, 12, and 24 h after hyperthermia. The cytoskeleton was observed with immunofluorescence. Flow cytometry and Western blotting were used to detect the expression of F-actin and relevant pathway protein. RESULTS: DnaJA4-KO and hyperthermia changed the cytoskeleton morphology of HaCaT cells. F-actin expression levels were elevated in DnaJA4-KO cells compared with WT cells (6364.33 ±â€Š989.10 vs. 4272.67 ±â€Š918.50, P < 0.05). In response to hyperthermia, F-actin expression levels of both WT and DnaJA4-KO cells showed a tendency to decrease followed by an obvious recovery after hyperthermia (WT cells: unheated vs. 6 h after hyperthermia or 24 h after hyperthermia: 0.34 ±â€Š0.02 vs. 0.24 ±â€Š0.01, 0.31 ±â€Š0.01, P < 0.001, P < 0.05; DnaJA4-KO cells: unheated vs. 6 h after hyperthermia or 24 h after hyperthermia: 0.44 ±â€Š0.01 vs. 0.30 ±â€Š0.01, 0.51 ±â€Š0.02, P < 0.001, P < 0.01). WT cells restored to baseline levels observed in the unheated condition, while DnaJA4-KO cells exceeded baseline levels in the recovery. As the upstream factors of F-actin, a similar profile in rho-associated serine/threonine kinase 1 (ROCK 1) and RhoA expressions was observed after hyperthermia. While E-cadherin expression was decreased in response to hyperthermia, it was increased in DnaJA4-KO cells compared with WT cells. CONCLUSIONS: Hyperthermia affects the expression levels of F-actin in HaCaT cells. DnaJA4 knockout increases the expression of F-actin in HaCaT cells after hyperthermia. DnaJA4 regulates the expressions of F-actin and the related pathway proteins in response to hyperthermia in HaCaT cells.

IL­18 knockout alleviates atopic dermatitis­like skin lesions induced by MC903 in a mouse model.

Interleukin (IL)­18, a pro­inflammatory cytokine, plays an important role in a number of skin diseases. The aim of the present study was to investigate the role of IL­18 in the development of atopic dermatitis (AD). For this purpose, mice were divided into 4 groups (n=5/group) as follows: i) The wild­type (WT) controls; ii) IL18 knockout (KO) controls; iii) MC903­treated WT mice; and iv) MC903­treated KO mice. MC903 (4 nmol in ethanol) was topically applied daily for 15 consecutive days to the exposed skins of mice. AD­like symptoms and severity were evaluated by the scoring of AD (SCORAD). Serum immunoglobulin E (IgE) and thymic stromal lymphopoietin (TSLP) levels were determined with the use of an enzyme­linked immunosorbent assay. Immunohistochemistry was used to assess the expression of IL­1ß, signal transducer and activator of transcription (STAT)3 and filaggrin (FLG) in the skin lesions. RT­qPCR was performed to assess the mRNA levels of IL­1ß, IL­4, IL­9, STAT3, corticotropin­releasing hormone receptor (CRHR)1, CRHR2, TSLP and caspase­1 in the skin lesions. It was demonstrated that IL­18 may function as a pleiotropic pro­inflammatory cytokine in the development of AD­like lesions. IL­18 KO reduced aggravated AD­like lesions induced by MC903, in part by upregulating Th2 cytokines. IL­18 promoted the expression of FLG in the epidermis and CRHR2 in AD­like lesions, but downregulated the serum levels of IgE. On the whole, the findings of the present study demonstrate that IL­18 deficiency alleviates AD­induced lesions in mice.

Association of positive and negative autologous serum skin test responses with clinical features of chronic spontaneous urticaria in Asian patients: A systematic review and meta-analysis.

Previous studies on the correlation between positive autologous serum skin test (ASST) responses and the clinical features of patients with chronic spontaneous urticaria (CSU) have provided conflicting results. To evaluate the significance of ASST responses in CSU, a variety of databases were searched from inception to March 2018 to identify relevant studies on CSU. Data were analyzed with use of the Cochrane Collaboration's Review Manager 5.2. Multiple relevant factors of CSU were evaluated by calculating the weighted mean difference, odds ratio and 95% confidence interval. The results indicated that CSU cases with positive ASST responses had higher urticaria activity scores and higher levels of total serum immunoglobulin E than CSU cases with negative responses in the ASST. In addition, a positive ASST response was more likely to be accompanied with the presence of thyroid autoantibodies and angioedema. An increased prevalence of CSU was identified in females, who were more likely to have a positive response in the ASST. It was also indicated that a greater incidence of positive ASST responses was present in CSU patients as compared with that in healthy controls. No statistically significant differences were obtained between positive and negative ASST responses with regard to age and duration of disease. Based on these results, it was concluded that the ASST provides an effective means of predicting urticaria activity and recurrence in CSU patients.

Interleukin-18 exacerbates skin inflammation and affects microabscesses and scale formation in a mouse model of imiquimod-induced psoriasis.

BACKGROUND: As a potent pro-inflammatory cytokine of the interleukin (IL)-1 family, IL-18 was elevated in early active and progressive plaque-type psoriatic lesions and that serum or plasma levels of IL-18 correlated with the Psoriasis Area and Severity Index (PASI). Although results from previous studies have established that IL-18 may aggravate psoriatic inflammation, the mechanisms of this process remain unknown. In this study, IL-18 knock out (KO) mice and wild-type (WT) mice were used to investigate the effects of IL-18 within a mouse model of psoriasis. METHODS: WT and IL-18 KO mice were divided into four groups, including imiquimod (IMQ)-treated IL-18 KO group (n = 11) and WT group (n = 13) as well as their respectively gene-matched control mice (receiving vaseline; n = 12). PASI scores were used to evaluate psoriatic lesions in IMQ-treated mice. Pathological features and dermal cellular infiltration were investigated by hematoxylin and eosin staining. The levels of psoriasis-related cytokines including IL-23, IL-17, IL-12, IL-1ß, IFNγ, IL-15, IL-27, and IL-4 were tested by real-time polymerase chain reaction (PCR). The protein level of IL-1ß, IL-27, CXCL1, and Ly6 g were investigated by immunohistochemistry (IHC). RESULTS: Acanthosis (98.46 ±â€Š14.12 vs. 222.68 ±â€Š71.10 µm, P < 0.01) and dermal cell infiltration (572.25 ±â€Š47.45 vs. 762.47 ±â€Š59.59 cells/field, P < 0.01) were significantly milder in IMQ-induced IL-18 KO mice compared with that in WT mice. IMQ-induced IL-18 KO mice manifested larger areas of Munro microabscesses (11,467.83 ±â€Š5112.09 vs. 4093.19 ±â€Š2591.88 µm, P < 0.01) and scales (100,935.24 ±â€Š41,167.77 vs. 41,604.41 ±â€Š14,184.10 µm, P < 0.01) as compared with WT mice. In skin lesions of IL-18 KO mice, the expressions of IL-1ß, IL-4, and IL-27 were all significantly upregulated but IL-17 was decreased. Histologically, strong positive signals of Ly6g were observed within the epidermis of IL-18 KO mice but expressions of CXCL1 were decreased. CONCLUSIONS: IL-18 may exacerbate prominent inflammation and influence pathological features in IMQ-induced mouse model of psoriasis. IL-18 may upregulate pro-inflammatory cytokines and reduce protective cytokines, thus aggravating psoriatic inflammation. In addition, IL-18 may be involved in the formation of Munro microabscesses and scales.

Local hyperthermia cleared multifarious viral warts in a patient with Cushing's syndrome.

A female Cushing's syndrome patient had been suffering from extensive viral warts for months. She was diagnosed with flat warts, common warts and plantar warts. The plantar warts on her right foot were initially treated using local hyperthermia at 44°C for 30 min according to a defined protocol, followed by treatment targeting a common wart on her left thumb. In response to hyperthermia, the flat warts on her eyelid dissipated within 12 weeks, and when combined with a 1 week administration of imiquimod, the common warts and plantar warts completely disappeared within 8 weeks. There were no signs of recurrence and during this treatment her Cushing's syndrome was alleviated. This pioneer trial suggests that local hyperthermia may serve as an effective mean for treating multiple cutaneous warts under the conditions of a systemic immuno-compromised disease.