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We explored the effect of a novel synthetic triterpenoid compound cyano enone of methyl boswellates (CEMB) on various prostate cancer and glioma cancer cell lines. CEMB displayed concentration-dependent cytotoxic activity with submicromolar lethal dose 50% (LD50) values in 10 of 10 tumor cell lines tested. CEMB-induced cytotoxicity is accompanied by activation of downstream effector caspases (caspases 3 and 7) and by upstream initiator caspases involved in both the extrinsic (caspase 8) and intrinsic (caspase 9) apoptotic pathways. By using short interfering RNAs (siRNA), we show evidence that knockdown of caspase 8, DR4, Apaf-1, and Bid impairs CEMB-induced cell death. Similar to other proapoptotic synthetic triterpenoid compounds, CEMB-induced apoptosis involved endoplasmic reticulum stress, as shown by partial rescue of tumor cells by siRNA-mediated knockdown of expression of genes involved in the unfolded protein response such as IRE1α, PERK, and ATF6. Altogether, our results suggest that CEMB stimulates several apoptotic pathways in cancer cells, suggesting that this compound should be evaluated further as a potential agent for cancer therapy. Mol Cancer Ther; 10(9); 1635–43. ©2011 AACR.

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Purpose:Radiomics is the extraction of multidimensional imaging features, which when correlated with genomics, is termed radiogenomics. However, radiogenomic biological validation is not sufficiently described in the literature. We seek to establish causality between differential gene expression status and MRI-extracted radiomic-features in glioblastoma.Experimental Design:Radiogenomic predictions and validation were done using the Cancer Genome Atlas and Repository of Molecular Brain Neoplasia Data glioblastoma patients (n = 93) and orthotopic xenografts (OX; n = 40). Tumor phenotypes were segmented, and radiomic-features extracted using the developed radiome-sequencing pipeline. Patients and animals were dichotomized on the basis of Periostin (POSTN) expression levels. RNA and protein levels confirmed RNAi-mediated POSTN knockdown in OX. Total RNA of tumor cells isolated from mouse brains (knockdown and control) was used for microarray-based expression profiling. Radiomic-features were utilized to predict POSTN expression status in patient, mouse, and interspecies.Results:Our robust pipeline consists of segmentation, radiomic-feature extraction, feature normalization/selection, and predictive modeling. The combination of skull stripping, brain-tissue focused normalization, and patient-specific normalization are unique to this study, providing comparable cross-platform, cross-institution radiomic features. POSTN expression status was not associated with qualitative or volumetric MRI parameters. Radiomic features significantly predicted POSTN expression status in patients (AUC: 76.56%; sensitivity/specificity: 73.91/78.26%) and OX (AUC: 92.26%; sensitivity/specificity: 92.86%/91.67%). Furthermore, radiomic features in OX were significantly associated with patients with similar POSTN expression levels (AUC: 93.36%; sensitivity/specificity: 82.61%/95.74%; P = 02.021E−15).Conclusions:We determined causality between radiomic texture features and POSTN expression levels in a preclinical model with clinical validation. Our biologically validated radiomic pipeline also showed the potential application for human–mouse matched coclinical trials.

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Melanin is usually formed by oxidative polymerization of phenolic compounds produced by bacteria, fungi, plants as well as animals. They have gained popularity for its multifarous properties and applications in various industries, especially the derma-cosmetic industries where fungal melanin has recently been highlighted as a potential source of natural bioactive compounds beneficial to skin and hair. The anti-oxidant potential and free radical scavenging activities of natural melanin makes them suitable for application in cosmetic products. In the present we have isolated and characterized a melanin pigment from Trichomerium bhatii NFCCI 4305, an unconventional fungus of Indian origin. The biological properties of purified melanin from the fungus were evaluated for its antioxidant activity. Following alkali acid treatment of the fermented broth, the pigment was extracted and further, its nature was confirmed as melanin through physico-chemical tests and spectral (UV and FTIR) analysis. The analysis of the elemental composition indicated that the pigment fall into DOPA category of melanin. The purified melanin showed a good DPPH (1, 1-Diphenyl-2-picrylhydrazyl) free radical-scavenging activity. To our understanding this is the first report on melanin characterization from the fungal genus Trichomerium.

This article is the 13th contribution in the Fungal Diversity Notes series, wherein 125 taxa from four phyla, ten classes, 31 orders, 69 families, 92 genera and three genera incertae sedis are treated, demonstrating worldwide and geographic distribution. Fungal taxa described and illustrated in the present study include three new genera, 69 new species, one new combination, one reference specimen and 51 new records on new hosts and new geographical distributions. Three new genera, Cylindrotorula (Torulaceae), Scolecoleotia (Leotiales genus incertae sedis) and Xenovaginatispora (Lindomycetaceae) are introduced based on distinct phylogenetic lineages and unique morphologies. Newly described species are Aspergillus lannaensis, Cercophora dulciaquae, Cladophialophora aquatica, Coprinellus punjabensis, Cortinarius alutarius, C. mammillatus, C. quercoflocculosus, Coryneum fagi, Cruentomycena uttarakhandina, Cryptocoryneum rosae, Cyathus uniperidiolus, Cylindrotorula indica, Diaporthe chamaeropicola, Didymella azollae, Diplodia alanphillipsii, Dothiora coronicola, Efibula rodriguezarmasiae, Erysiphe salicicola, Fusarium queenslandicum, Geastrum gorgonicum, G. hansagiense, Helicosporium sexualis, Helminthosporium chiangraiensis, Hongkongmyces kokensis, Hydrophilomyces hydraenae, Hygrocybe boertmannii, Hyphoderma australosetigerum, Hyphodontia yunnanensis, Khaleijomyces umikazeana, Laboulbenia divisa, Laboulbenia triarthronis, Laccaria populina, Lactarius pallidozonarius, Lepidosphaeria strobelii, Longipedicellata megafusiformis, Lophiotrema lincangensis, Marasmius benghalensis, M. jinfoshanensis, M. subtropicus, Mariannaea camelliae, Melanographium smilaxii, Microbotryum polycnemoides, Mimeomyces digitatus, Minutisphaera thailandensis, Mortierella solitaria, Mucor harpali, Nigrograna jinghongensis, Odontia huanrenensis, O. parvispina, Paraconiothyrium ajrekarii, Parafuscosporella niloticus, Phaeocytostroma yomensis, Phaeoisaria synnematicus, Phanerochaete hainanensis, Pleopunctum thailandicum, Pleurotheciella dimorphospora, Pseudochaetosphaeronema chiangraiense, Pseudodactylaria albicolonia, Rhexoacrodictys nigrospora, Russula paravioleipes, Scolecoleotia eriocamporesi, Seriascoma honghense, Synandromyces makranczyi, Thyridaria aureobrunnea, Torula lancangjiangensis, Tubeufia longihelicospora, Wicklowia fusiformispora, Xenovaginatispora phichaiensis and Xylaria apiospora. One new combination, Pseudobactrodesmium stilboideus is proposed. A reference specimen of Comoclathris permunda is designated. New host or distribution records are provided for Acrocalymma fici, Aliquandostipite khaoyaiensis, Camarosporidiella laburni, Canalisporium caribense, Chaetoscutula juniperi, Chlorophyllum demangei, C. globosum, C. hortense, Cladophialophora abundans, Dendryphion hydei, Diaporthe foeniculina, D. pseudophoenicicola, D. pyracanthae, Dictyosporium pandanicola, Dyfrolomyces distoseptatus, Ernakulamia tanakae, Eutypa flavovirens, E. lata, Favolus septatus, Fusarium atrovinosum, F. clavum, Helicosporium luteosporum, Hermatomyces nabanheensis, Hermatomyces sphaericoides, Longipedicellata aquatica, Lophiostoma caudata, L. clematidis-vitalbae, Lophiotrema hydei, L. neoarundinaria, Marasmiellus palmivorus, Megacapitula villosa, Micropsalliota globocystis, M. gracilis, Montagnula thailandica, Neohelicosporium irregulare, N. parisporum, Paradictyoarthrinium diffractum, Phaeoisaria aquatica, Poaceascoma taiwanense, Saproamanita manicata, Spegazzinia camelliae, Submersispora variabilis, Thyronectria caudata, T. mackenziei, Tubeufia chiangmaiensis, T. roseohelicospora, Vaginatispora nypae, Wicklowia submersa, Xanthagaricus necopinatus and Xylaria haemorrhoidalis. The data presented herein are based on morphological examination of fresh specimens, coupled with analysis of phylogenetic sequence data to better integrate taxa into appropriate taxonomic ranks and infer their evolutionary relationships.

BackgroundThe aim of this study was to describe the health-seeking journey for patients with microbial keratitis (MK) in Nepal and identify factors associated with delay.MethodsProspective cohort study where MK patients attending a large, tertiary-referral eye hospital in south-eastern Nepal between June 2019 and November 2020 were recruited. We collected demographic details, clinical history, and examination findings. Care-seeking journey details were captured including places attended, number of journeys, time from symptom onset, and costs. We compared “direct” with “indirect” presenters, analyzing for predictors of delay.ResultsWe enrolled 643 patients with MK. The majority (96%) self-referred. “Direct” attenders accounted for only 23.6% (152/643) of patients, the majority of “indirect” patients initially presented to a pharmacy (255/491). Over half (328/643) of all cases presented after at least 7 days. The total cost of care increased with increasing numbers of facilities visited (p < 0.001). Those living furthest away were least likely to present directly (p < 0.001). Factors independently associated with delayed presentation included distance >50 km from the eye hospital [aOR 5.760 (95% CI 1.829–18.14, p = 0.003)], previous antifungal use [aOR 4.706 (95% CI 3.139–5.360)], and two or more previous journeys [aOR 1.442 (95% CI 1.111–3.255)].ConclusionsMost patients visited at least one facility prior to our institution, with time to presentation and costs increasing with the number of prior journeys. Distance to the eye hospital is a significant barrier to prompt, direct presentation. Based on these findings, improving access to eye care services, strengthening referral networks and encouraging early appropriate treatment are recommended to reduce delay, ultimately improving clinical outcomes.

Background The tomato leafminer, Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae), is a serious pest of solanaceous plant species, mainly Solanum lycopersicum L. The entomopathogenic fungi, an alternative to chemical insecticides, proved to be an efficient biocontrol agent in reducing pest population density. In the present study, an entomopathogenic fungus, Purpureocillium lilacinum isolated from soil was identified based on the morphological and molecular characteristics and its pathogenicity was tested against target pest. Results The micromorphological characters showed variations in growth pattern, shape and colour on different cultural media. For molecular analysis, a phylogenetic tree based on ITS/LSU and ITS/β-tubulin (benA) gene regions was constructed which revealed the isolate (FC18) as P. lilacinum (Thom) Luangsa-ard, Houbraken, Hywel-Jones and Samson (Family: Ophiocordycipitaceae). Further, the pathogenicity of P. lilacinum was tested using different spore concentrations (1 × 108, 1 × 107, 1 × 106, 1 × 105 and 1 × 104 spores/ml) on larval and pupal stages of T. absoluta, which showed a dose-dependent mortality. At the highest concentration (1 × 108 spores/ml), the mean mortality of 92.99, 83.05, 72.0% of second, third and fourth instar was observed, respectively. Pupal mortality also showed significant differences at different spore concentrations. Conclusion Obtained results showed that the virulence of the indigenous strain of P. lilacinum on T. absoluta can be utilized in the field suppression of the pest as a potent biocontrol agent.

Background Oncolytic adenoviruses are promising new treatments against solid tumors, particularly for glioblastoma (GBM), and preclinical models are required to evaluate the mechanisms of efficacy. However, due to the species selectivity of adenovirus, there is currently no single animal model that supports viral replication, tumor oncolysis, and a virus-mediated immune response. To address this gap, we took advantage of the Syrian hamster to develop the first intracranial glioma model that is both adenovirus replication-permissive and immunocompetent. Methods We generated hamster glioma stem-like cells (hamGSCs) by transforming hamster neural stem cells with hTERT, simian virus 40 large T antigen, and h-RasV12. Using a guide-screw system, we generated an intracranial tumor model in the hamster. The efficacy of the oncolytic adenovirus Delta-24-RGD was assessed by survival studies, and tumor-infiltrating lymphocytes (TILs) were evaluated by flow cytometry. Results In vitro, hamGSCs supported viral replication and were susceptible to Delta-24-RGD mediated cell death. In vivo, hamGSCs consistently developed into highly proliferative tumors resembling high-grade glioma. Flow cytometric analysis of hamster gliomas revealed significantly increased T-cell infiltration in Delta-24-RGD infected tumors, indicative of immune activation. Treating tumor-bearing hamsters with Delta-24-RGD led to significantly increased survival compared to hamsters treated with phosphate buffered saline (PBS). Conclusions This adenovirus-permissive, immunocompetent hamster glioma model overcomes the limitations of previous model systems and provides a novel platform to study the interactions between tumor cells, the host immune system, and oncolytic adenoviral therapy; understanding of which will be critical to implementing oncolytic adenovirus in the clinic.

Neutrophil-macrophage interplay is a fine-tuning mechanism that regulates the innate immune response during infection and inflammation. Cell surface receptors play an essential role in neutrophil and macrophage functions. The same receptor can provide different outcomes within diverse leukocyte subsets in different inflammatory conditions. Understanding the variety of responses mediated by one receptor is critical for the development of anti-inflammatory treatments. In this study, we evaluated the role of a leukocyte adhesive receptor, integrin αDβ2, in the development of acute inflammation. αDβ2 is mostly expressed on macrophages and contributes to the development of chronic inflammation. In contrast, we found that αD-knockout dramatically increases mortality in the cecal ligation and puncture sepsis model and LPS-induced endotoxemia. This pathologic outcome of αD-deficient mice is associated with a reduced number of monocyte-derived macrophages and an increased number of neutrophils in their lungs. However, the tracking of adoptively transferred fluorescently labeled wild-type (WT) and αD−/− monocytes in WT mice during endotoxemia demonstrated only a moderate difference between the recruitment of these two subsets. Moreover, the rescue experiment, using i.v. injection of WT monocytes to αD-deficient mice followed by LPS challenge, showed only slightly reduced mortality. Surprisingly, the injection of WT neutrophils to the bloodstream of αD−/− mice markedly increased migration of monocyte-derived macrophage to lungs and dramatically improves survival. αD-deficient neutrophils demonstrate increased necrosis/pyroptosis. αDβ2-mediated macrophage accumulation in the lungs promotes efferocytosis that reduced mortality. Hence, integrin αDβ2 implements a complex defense mechanism during endotoxemia, which is mediated by macrophages via a neutrophil-dependent pathway.

BackgroundOncolytic viruses are considered part of immunotherapy and have shown promise in preclinical experiments and clinical trials. Results from these studies have suggested that tumor microenvironment remodeling is required to achieve an effective response in solid tumors. Here, we assess the extent to which targeting specific mechanisms underlying the immunosuppressive tumor microenvironment optimizes viroimmunotherapy.MethodsWe used RNA-seq analyses to analyze the transcriptome, and validated the results using Q-PCR, flow cytometry, and immunofluorescence. Viral activity was analyzed by replication assays and viral titration. Kyn and Trp metabolite levels were quantified using liquid chromatography–mass spectrometry. Aryl hydrocarbon receptor (AhR) activation was analyzed by examination of promoter activity. Therapeutic efficacy was assessed by tumor histopathology and survival in syngeneic murine models of gliomas, including Indoleamine 2,3-dioxygenase (IDO)-/- mice. Flow cytometry was used for immunophenotyping and quantification of cell populations. Immune activation was examined in co-cultures of immune and cancer cells. T-cell depletion was used to identify the role played by specific cell populations. Rechallenge experiments were performed to identify the development of anti-tumor memory.ResultsBulk RNA-seq analyses showed the activation of the immunosuppressive IDO-kynurenine-AhR circuitry in response to Delta-24-RGDOX infection of tumors. To overcome the effect of this pivotal pathway, we combined Delta-24-RGDOX with clinically relevant IDO inhibitors. The combination therapy increased the frequency of CD8+ T cells and decreased the rate of myeloid-derived suppressor cell and immunosupressive Treg tumor populations in animal models of solid tumors. Functional studies demonstrated that IDO-blockade-dependent activation of immune cells against tumor antigens could be reversed by the oncometabolite kynurenine. The concurrent targeting of the effectors and suppressors of the tumor immune landscape significantly prolonged the survival in animal models of orthotopic gliomas.ConclusionsOur data identified for the first time the in vivo role of IDO-dependent immunosuppressive pathways in the resistance of solid tumors to oncolytic adenoviruses. Specifically, the IDO-Kyn-AhR activity was responsible for the resurface of local immunosuppression and resistance to therapy, which was ablated through IDO inhibition. Our data indicate that combined molecular and immune therapy may improve outcomes in human gliomas and other cancers treated with virotherapy.

Though soils sequester large amounts of carbon (C), variations in physical and chemical characteristics of top and deep layers necessitate the study of factors governing topsoil and deep soil C seq...

Arboviruses maintain high mutation rates due to lack of proofreading ability of their viral polymerases, in some cases facilitating adaptive evolution and emergence. Here we show that, just before its 2013 spread to the Americas, Zika virus (ZIKV) underwent an envelope protein V473M substitution (E-V473M) that increased neurovirulence, maternal-to-fetal transmission, and viremia to facilitate urban transmission. A preepidemic Asian ZIKV strain (FSS13025 isolated in Cambodia in 2010) engineered with the V473M substitution significantly increased neurovirulence in neonatal mice and produced higher viral loads in the placenta and fetal heads in pregnant mice. Conversely, an epidemic ZIKV strain (PRVABC59 isolated in Puerto Rico in 2015) engineered with the inverse M473V substitution reversed the pathogenic phenotypes. Although E-V473M did not affect oral infection of Aedes aegypti mosquitoes, competition experiments in cynomolgus macaques showed that this mutation increased its fitness for viremia generation, suggesting adaptive evolution for human viremia and hence transmission. Mechanistically, the V473M mutation, located at the second transmembrane helix of the E protein, enhances virion morphogenesis. Overall, our study revealed E-V473M as a critical determinant for enhanced ZIKV virulence, intrauterine transmission during pregnancy, and viremia to facilitate urban transmission.

Fungal diversity notes is one of the important journal series of fungal taxonomy that provide detailed descriptions and illustrations of new fungal taxa, as well as providing new information of fungal taxa worldwide. This article is the 11th contribution to the fungal diversity notes series, in which 126 taxa distributed in two phyla, six classes, 24 orders and 55 families are described and illustrated. Taxa in this study were mainly collected from Italy by Erio Camporesi and also collected from China, India and Thailand, as well as in some other European, North American and South American countries. Taxa described in the present study include two new families, 12 new genera, 82 new species, five new combinations and 25 new records on new hosts and new geographical distributions as well as sexual-asexual reports. The two new families areEriomycetaceae(Dothideomycetes, familyincertae sedis) andFasciatisporaceae(Xylariales, Sordariomycetes). The twelve new genera compriseBhagirathimyces(Phaeosphaeriaceae),Camporesiomyces(Tubeufiaceae),Eriocamporesia(Cryphonectriaceae),Eriomyces(Eriomycetaceae),Neomonodictys(Pleurotheciaceae),Paraloratospora(Phaeosphaeriaceae),Paramonodictys(Parabambusicolaceae),Pseudoconlarium(Diaporthomycetidae, genusincertae sedis),Pseudomurilentithecium(Lentitheciaceae),Setoapiospora(Muyocopronaceae),Srinivasanomyces(Vibrisseaceae) andXenoanthostomella(Xylariales, generaincertae sedis). The 82 new species compriseAcremonium chiangraiense,Adustochaete nivea,Angustimassarina camporesii,Bhagirathimyces himalayensis,Brunneoclavispora camporesii,Camarosporidiella camporesii,Camporesiomyces mali,Camposporium appendiculatum,Camposporium multiseptatum,Camposporium septatum,Canalisporium aquaticium,Clonostachys eriocamporesiana,Clonostachys eriocamporesii,Colletotrichum hederiicola,Coniochaeta vineae,Conioscypha verrucosa,Cortinarius ainsworthii,Cortinarius aurae,Cortinarius britannicus,Cortinarius heatherae,Cortinarius scoticus,Cortinarius subsaniosus,Cytospora fusispora,Cytospora rosigena,Diaporthe camporesii,Diaporthe nigra,Diatrypella yunnanensis,Dictyosporium muriformis,Didymella camporesii,Diutina bernali,Diutina sipiczkii,Eriocamporesia aurantia,Eriomyces heveae,Ernakulamia tanakae,Falciformispora uttaraditensis,Fasciatispora cocoes,Foliophoma camporesii,Fuscostagonospora camporesii,Helvella subtinta,Kalmusia erioi,Keissleriella camporesiana,Keissleriella camporesii,Lanspora cylindrospora,Loratospora arezzoensis,Mariannaea atlantica,Melanographium phoenicis,Montagnula camporesii,Neodidymelliopsis camporesii,Neokalmusia kunmingensis,Neoleptosporella camporesiana,Neomonodictys muriformis,Neomyrmecridium guizhouense,Neosetophoma camporesii,Paraloratospora camporesii,Paramonodictys solitarius,Periconia palmicola,Plenodomus triseptatus,Pseudocamarosporium camporesii,Pseudocercospora maetaengensis,Pseudochaetosphaeronema kunmingense,Pseudoconlarium punctiforme,Pseudodactylaria camporesiana,Pseudomurilentithecium camporesii,Pseudotetraploa rajmachiensis,Pseudotruncatella camporesii,Rhexocercosporidium senecionis,Rhytidhysteron camporesii,Rhytidhysteron erioi,Septoriella camporesii,Setoapiospora thailandica,Srinivasanomyces kangrensis,Tetraploa dwibahubeeja,Tetraploa pseudoaristata,Tetraploa thrayabahubeeja,Torula camporesii,Tremateia camporesii,Tremateia lamiacearum,Uzbekistanica pruni,Verruconis mangrovei,Wilcoxina verruculosa,Xenoanthostomella chromolaenaeandXenodidymella camporesii. The five new combinations areCamporesiomyces patagoniensis,Camporesiomyces vaccinia,Camposporium lycopodiellae,Paraloratospora gahniaeandRhexocercosporidium microsporum. The 22 new records on host and geographical distribution compriseArthrinium marii,Ascochyta medicaginicola,Ascochyta pisi,Astrocystis bambusicola,Camposporium pellucidum,Dendryphiella phitsanulokensis,Diaporthe foeniculina,Didymella macrostoma,Diplodia mutila,Diplodia seriata,Heterosphaeria patella,Hysterobrevium constrictum,Neodidymelliopsis ranunculi,Neovaginatispora fuckelii,Nothophoma quercina,Occultibambusa bambusae,Phaeosphaeria chinensis,Pseudopestalotiopsis theae,Pyxine berteriana,Tetraploa sasicola,Torula gaodangensisandWojnowiciella dactylidis. In addition, the sexual morphs ofDissoconium eucalyptiandPhaeosphaeriopsis pseudoagavacearumare reported fromLaurus nobilisandYucca gloriosain Italy, respectively. The holomorph ofDiaporthe cynaroidisis also reported for the first time.

Filamentous fungi synthesize natural products as an ecological function. In this study, an interesting indigenous fungus producing orange pigment exogenously was investigated in detail as it possesses additional attributes along with colouring properties. An interesting fungus was isolated from a dicot plant, Maytenus rothiana. After a detailed study, the fungal isolate turned out to be a species of Gonatophragmium belonging to the family Acrospermaceae. Based on the morphological, cultural, and sequence-based phylogenetic analysis, the identity of this fungus was confirmed as Gonatophragmium triuniae. Although this fungus grows moderately, it produces good amounts of pigment on an agar medium. The fermented crude extract isolated from G. triuniae has shown antioxidant activity with an IC50 value of 0.99 mg/mL and antibacterial activity against Gram-positive bacteria (with MIC of 3.91 μg/mL against Bacillus subtilis, and 15.6 μg/mL and 31.25 μg/mL for Staphylococcus aureus and Micrococcus luteus, respectively). Dyeing of cotton fabric mordanted with FeSO4 using crude pigment was found to be satisfactory based on visual observation, suggesting its possible use in the textile industry. The orange pigment was purified from the crude extract by preparative HP-TLC. In addition, UV-Vis, FTIR, HRMS and NMR (1H NMR, 13C NMR), COSY, and DEPT analyses revealed the orange pigment to be “1,2-dimethoxy-3H-phenoxazin-3-one” (C14H11NO4, m/z 257). To our understanding, the present study is the first comprehensive report on Gonatophragmium triuniae as a potential pigment producer, reporting “1,2-dimethoxy-3H-phenoxazin-3-one” as the main pigment from the crude hexane extract. Moreover, this is the first study reporting antioxidant, antibacterial, and dyeing potential of crude extract of G. triuniae, suggesting possible potential applications of pigments and other bioactive secondary metabolites of the G. triuniae in textile and pharmaceutical industry.

Sunflower (Helianthus annuus) is one of the most important oilseed crops worldwide. However, the transcriptional regulation underlying oil accumulation in sunflower is not fully understood. WRINKLED1 (WRI1) is an essential transcription factor governing oil accumulation in plant cells. Here, we identify and characterize a sunflower ortholog of WRI1 (HaWRI1), which is highly expressed in developing seeds. Transient production of HaWRI1 stimulated substantial oil accumulation in Nicotiana benthamiana leaves. Dual-luciferase reporter assay, electrophoretic mobility shift assay, fatty acid quantification, and gene expression analysis demonstrate that HaWRI1 acts as a pivotal transcription factor controlling the expression of genes involved in late glycolysis and fatty acid biosynthesis. HaWRI1 directly binds to the cis-element, AW-box, in the promoter of biotin carboxyl carrier protein isoform 2 (BCCP2). In addition, we characterize an 80 amino-acid C-terminal domain of HaWRI1 that is crucial for transactivation. Moreover, seed-specific overexpression of HaWRI1 in Arabidopsis plants leads to enhanced seed oil content as well as upregulation of the genes involved in fatty acid biosynthesis. Taken together, our work demonstrates that HaWRI1 plays a pivotal role in the transcriptional control of seed oil accumulation, providing a potential target for bioengineering sunflower oil yield improvement. Ministry of Education (MOE) Published version This work was supported by the Ministry of Education (MOE) of Singapore Tier 1 to W.M. (grant number: RG29/20) and MOE of Singapore Tier 2 to W.M. (grant number: MOE-T2EP30220-0011).

In order to support livelihoods, enhance food security, restore ecosystem services, and reduce pressure on forests, degraded land can be restored by utilising alternative land-use systems (ALUS), such as silviculture, silvipasture, and hortipasture techniques. ALUS significantly modify the dynamics of soil nutrients in both the surface and subsurface layers. Soils from the 0–15, 15–30, and 30–45 cm layers of Leucaena leucocephala (S)-, Hardwickia binata (H)-, Emblica officinalis (A)-, and Azadiracta indica (N)-based silviculture systems, Acacia nilotica-based silvipasture systems (SPS), natural grassland (NT), and fallow land (F) were sampled in order to better understand the nutrient dynamics of ALUS. Soils under S, H, and SPS had ~203%, 195%, and 129% higher organic carbon (SOC), respectively, than fallow land in the 0–15 cm soil layer. In the subsequent soil layer, those land-use systems had ~199%, 82%, and 110% higher SOC, respectively, than fallow land. Similarly, in the deeper layer, those land uses had ~232%, 23%, and 105% higher SOC, respectively, than fallow land. SPS and NT also improved the SOC concentration significantly over fallow land. Plots under S, H, and SPS had ~198%, 190%, and 125% higher available N, respectively, than fallow land in 0–15 cm soil layer. In the 15–30 cm soil layer, those land-use systems had ~19%9, 82%, and 110% higher available N, respectively, than fallow land. These systems also improved the P and K contents in subsurface soil. Micronutrient concentrations were also improved in soils under S, H, and SPS. Hence, ALUS’ adoption in degraded areas with trees provides a chance for C storage and improves the nutrient dynamics on degraded land.

Filamentous fungi synthesize natural products as an ecological function. In this study, an interesting indigenous fungus producing orange pigment exogenously was investigated in detail as it possesses additional attributes along with colouring properties. An interesting fungus was isolated from a dicot plant, Maytenus rothiana. After a detailed study, the fungal isolate turned out to be a species of Gonatophragmium belonging to the family Acrospermaceae. Based on the morphological, cultural, and sequence-based phylogenetic analysis, the identity of this fungus was confirmed as Gonatophragmium triuniae. Although this fungus grows moderately, it produces good amounts of pigment on an agar medium. The fermented crude extract isolated from G. triuniae has shown antioxidant activity with an IC50 value of 0.99 mg/mL and antibacterial activity against Gram-positive bacteria (with MIC of 3.91 μg/mL against Bacillus subtilis, and 15.6 μg/mL and 31.25 μg/mL for Staphylococcus aureus and Micrococcus luteus, respectively). Dyeing of cotton fabric mordanted with FeSO4 using crude pigment was found to be satisfactory based on visual observation, suggesting its possible use in the textile industry. The orange pigment was purified from the crude extract by preparative HP-TLC. In addition, UV-Vis, FTIR, HRMS and NMR (1H NMR, 13C NMR), COSY, and DEPT analyses revealed the orange pigment to be “1,2-dimethoxy-3H-phenoxazin-3-one” (C14H11NO4, m/z 257). To our understanding, the present study is the first comprehensive report on Gonatophragmium triuniae as a potential pigment producer, reporting “1,2-dimethoxy-3H-phenoxazin-3-one” as the main pigment from the crude hexane extract. Moreover, this is the first study reporting antioxidant, antibacterial, and dyeing potential of crude extract of G. triuniae, suggesting possible potential applications of pigments and other bioactive secondary metabolites of the G. triuniae in textile and pharmaceutical industry.

Oncolytic viruses are considered part of immunotherapy and have shown promise in preclinical experiments and clinical trials in a small fraction of patients. To improve the response rate, it is necessary to reshape the tumor microenvironment that shields the tumor from the immune system of the patient. We engineered Delta-24-RGDOX (DNX-2440), an oncolytic adenovirus to express the T-cell activator OX40L, which triggered, in murine glioblastoma models, a dramatic reshaping of the tumor microenvironment dominated by changes in immune processes in an RNA-seq analysis of ingenuity pathways. Paradoxically, network analyses also revealed that Delta-24-RGDOX induced robust activation of the cytokine-driven immunosuppressive IDO-Kynurenine-AhR circuitry, indicating a potential mechanism of resistance of gliomas to oncolytic virotherapy. To reverse this immunosuppression, we combined Delta-24-RGDOX with clinically relevant IDO inhibitors to treat glioma-bearing mice. The combined therapy decreased the activation of the IDO network without affecting viral activity. Flow cytometry assays revealed a decrease in the presence of the immunosuppressive cell populations, MDSCs and Tregs, which presence was higher in Delta-24-RGDOX-treated tumors. These data were further confirmed in gene set enrichment analyses. Functional co-culture studies showed that the combined therapy activated splenocytes against tumor antigens and that this activation was reversed by Kynurenine. Importantly, the combination treatment eradicated the tumors in a CD4-dependent manner and significantly prolonged the survival of glioma-bearing mice. Altogether, these studies indicate that the combination treatment promotes an adaptive immune response while decreasing the immunosuppression caused by the virus-induced IDO activation. Furthermore, our data identified the striking role of immunosuppressive pathways in the resistance of gliomas to oncolytic virotherapy. Specifically, the activity of the tumor microenvironment IDO circuitry was responsible, at least partially, for the remodeling of local immunosuppression after tumor infection. Combining molecular and immune-related therapies may improve outcomes in human gliomas treated with virotherapy.