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Unearthing the fungal endophyte Aspergillus terreus for chemodiversity and medicinal prospects: a comprehensive review

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Fungal Biology and Biotechnology 2023年第1期10卷 1页

作者： Amr, Khadiga Ibrahim, Nehal Elissawy, Ahmed M. Singab, Abdel Nasser B. Department of Pharmacognosy Faculty of Pharmacy Ain-Shams University Organization of African Unity Street 1 Cairo 11566 Egypt Center of Drug Discovery Research and Development Ain-Shams University Organization of African Unity Street 1 Cairo 11566 Egypt

Aspergillus terreus microorganism represents a promising prospective source for drug discovery since it is rich in diverse kinds of bioactive secondary metabolites. It contributed to many biotechnological applications and its metabolites are used in the synthesis of certain pharmaceuticals and food products, in addition to its useful uses in fermentation processes. There are about 346 compounds identified from marine and terrestrial-derived A. terreus from 1987 until 2022, 172 compounds of them proved a vast array of bioactivity. This review aimed to create an up-to-date comprehensive literature data of A. terreus’s secondary metabolites classes supported by its different bioactivity data to be a scientific record for the next work in drug discovery. © 2023, The Author(s).

关键词： A. terreus Biological activity Drug discovery Fungal endophytes Natural products

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development of an Automated Method for Compiling a List of Protected Objects Based on a Special Classification of Information Assets in the Field of Information Security

Development of an Automated Method for Compiling a List of P...

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2023 International Russian Smart Industry Conference, SmartIndustryCon 2023

作者： Manzhosov, A.V. Bolodurina, I.P. Dolgushev, N.A. Research Center Unity in Development Ekaterinburg Russia Orenburg State University Department of Applied Mathematics Orenburg Russia Orenburg State University Department of Computer Engineering and Information Security Orenburg Russia

ISBN: (纸本)9781665464291

This article reviewed the sources on the research topic, as a result of which it was determined that the research topic is discussed, relevant in the Russian and international agenda. A comparison of asset classifications in legal acts was carried out, which determines the recommended basic list of assets. A special classification of information assets has been developed, which systematizes the classes of financial assets according to the criterion of the possibility of information security risks affecting an information asset. An algorithm for the rational construction of a list of financial assets in the field of information security has been developed. An automated tool has been developed to determine the totality of assets based on special allocations. © 2023 IEEE.

关键词： Classification (of information)

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development of an Automated Method for Compiling a List of Protected Objects Based on a Special Classification of Information Assets in the Field of Information Security

Development of an Automated Method for Compiling a List of P...

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International Russian Smart Industry Conference (SmartIndustryCon)

作者： A. V. Manzhosov I. P. Bolodurina N. A. Dolgushev Research Center Unity in Development Ekaterinburg Russia Department of Applied Mathematics Orenburg State University Orenburg Russia Department of Computer Engineering and Information Security Orenburg State University Orenburg Russia

关键词： Industries Law Information security Classification algorithms Resource management Task analysis

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Discovery of Novel Zoanthamine-Type Alkaloids Derived from Cultured Zoanthus Kuroshio with Prominent Therapeutic Potential Against Osteoporosis

SSRN

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SSRN 2025年

作者： Pham, Ngoc-Thac Peng, Bo-Rong Le, Huong-Giang Chen, Yun-Shiuan Huynh, Thanh Hao Chen, Lo-Yun Nguyen, Le Anh Tuan Nguyen, Dang T. Su, Jui-Hsin El-Shazly, Mohamed Lee, Mei-Hsien Lai, Kuei-Hung PhD Program in Clinical Drug Development of Herbal Medicine College of Pharmacy Taipei Medical University Taipei110301 Taiwan Graduate Institute of Pharmacognosy College of Pharmacy Taipei Medical University Taipei110301 Taiwan Graduate Institute of Healthy Industry Technology Research Center for Chinese Herbal Medicine College of Human Ecology Chang Gung University of Science and Technology Taoyuan333324 Taiwan Faculty of Pharmacy Lac Hong University Dongnai Bien Hoa Viet Nam Faculty of Applied Sciences Ton Duc Thang University Ho Chi Minh City Viet Nam National Museum of Marine Biology and Aquarium Pingtung944401 Taiwan Department of Marine Biotechnology and Resources National Sun Yat-sen University Kaohsiung804201 Taiwan Department of Pharmacognosy Faculty of Pharmacy Ain-Shams University Organization of African Unity Street Abassia Cairo11566 Egypt Center for Reproductive Medicine and Sciences Taipei Medical University Hospital Taipei110301 Taiwan Traditional Herbal Medicine Research Center Taipei Medical University Hospital Taipei110301 Taiwan

Marine invertebrates are well-established as prolific sources of secondary metabolites, renowned for their remarkable chemical diversity and extensive spectrum of biological activities. Among these, Zoanthus Kuroshio (ZK) has gained significant attention due to its abundant chemical constituents, particularly alkaloids, which exhibit promising bioactivities against osteoporosis. To investigate its therapeutic potential, the extracts of cultured ZK exhibited significant anti-osteoporotic activity in vitro with MG-63 cells, leading to the integration of Global Natural Products Social Molecular Networking (GNPS) to identify fractions containing the highest amounts of alkaloids. Subsequent isolation efforts resulted in the discovery of five novel zoanthamine-type alkaloids: norzoabenzaldehyde (1), norzoazepanol (2), 11-keto-3-acetoxynorzoanthamine (3), 11-hydroxynorzoanthamide B (4), and 11-hydroxyzoanthamide B (5), alongside four previously identified compounds, including norzoanthaminone (6), 3-acetoxynorzoanthamine (7), 3-acetoxyzoanthamine (8), and 3-hydroxynorzoanthamine (9). Structural elucidation of compounds 1, 3, 4, and 5 was achieved using comprehensive spectroscopic analyses, whereas compound 2 required advanced techniques, including time-dependent density functional theory nuclear magnetic resonance (TDDFT-NMR) calculations and comparative analysis of theoretical and experimental electronic circular dichroism (ECD) data, to ascertain its absolute configuration. Bioassays evaluating alkaline phosphatase activity and mineralization demonstrated that compound 9 exhibited significant activity, with an IC50 value of 20 µM. The mechanism by which this compound operates was elucidated through network pharmacology, molecular docking and dynamics simulations, further confirming the stability of interactions between compound 9 and key protein targets, thereby emphasizing its potential as a promising lead compound for osteoporosis therapeutics. © 2025, The Authors. All

关键词： Spectroscopic analysis

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Application of response surface methodology and quantitative NMR for the optimum extraction, characterization, and quantitation of Antrodia cinnamomea triterpenoids

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Scientific Reports 2023年第1期13卷 1-13页

作者： Li, Chi-Ying Wu, Yang-Chang Chang, Fang-Rong El-Shazly, Mohamed Du, Ying-Chi Lu, Chi-Yu Duh, Tsai-Hui Wu, Tung-Ying Graduate Institute of Natural Products College of Pharmacy Kaohsiung Medical University Kaohsiung 807 Taiwan Department of Pharmacology and Pharmaceutical Sciences School of Pharmacy University of Southern California 1985 Zonal Avenue Los Angeles 90089 CA United States Graduate Institute of Integrated Medicine China Medical University Taichung 404 Taiwan Chinese Medicine Research and Development Center China Medical University Hospital Taichung 404 Taiwan Drug Development and Value Creation Research Center Kaohsiung Medical University Kaohsiung 807 Taiwan Department of Medical Research Kaohsiung Medical University Hospital Kaohsiung Medical University Kaohsiung 807 Taiwan Department of Marine Biotechnology and Resources National Sun Yat-sen University Kaohsiung 804 Taiwan Department of Pharmacognosy Faculty of Pharmacy Ain-Shams University Organization of African Unity Street Abassia Cairo 11566 Egypt Department of Biochemistry College of Medicine Kaohsiung Medical University Kaohsiung 807 Taiwan Department of Medicinal and Applied Chemistry Kaohsiung Medical University Kaohsiung 807 Taiwan Research Center for Precision Environmental Medicine Kaohsiung Medical University Kaohsiung 807 Taiwan Department of Biological Science and Technology Meiho University Pingtung 912 Taiwan Department of Food Science and Nutrition Meiho University Pingtung 912 Taiwan

Antrodia cinnamomea (AC) is a treasured Asian medicinal mushroom, which has attracted attention due to recent research on its effectiveness in targeting a variety of serious ailments such as cancer and liver diseases. Among different A. cinnamomea constituents, triterpenoids are regarded as the most therapeutically attractive components because of their anti-inflammatory and cytotoxic activities. In the present study, we proposed a mathematical and statistical extraction protocol to evaluate the concentrations of total ergostane and lanostane triterpenoid derivatives from the ethanolic extract of the wild fruiting bodies of A. cinnamomea (EEAC) by utilizing response surface methodology (RSM) and quantitative NMR (qNMR) approaches. The optimum response surface model showed that the variations of the investigated response variables reached more than 90%, suggesting that the developed model is accurate in explaining response variability. Furthermore, the EEAC major characteristic triterpenoids were quantified through the comparison of the HPLC-tandem MS results with those of the qNMR results. The precision of the used techniques was also evaluated. The experimental design of the EEAC optimum extraction procedure obtained by using RSM and qNMR enabled accurate characterization and quantitation of A. cinnamomea triterpenoids. © 2023, The Author(s).

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Unprecedented anti-inflammatory biscembranoids with 3-hydroperoxy-3-methylcyclohex-1-ene moiety isolated from aquaculture Sarcophyton trocheliophorum

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Aquaculture Reports 2023年 33卷

作者： Peng, Bo-Rong Nguyen, Ngoc Bao An Chen, Lo-Yun El-Shazly, Mohamed Hwang, Tsong-Long Su, Jui-Hsin Lai, Kuei-Hung Graduate Institute of Pharmacognosy College of Pharmacy Taipei Medical University Taipei 11031 Taiwan PhD Program in Clinical Drug Development of Herbal Medicine College of Pharmacy Taipei Medical University Taipei 11031 Taiwan Department of Pharmacognosy Faculty of Pharmacy Ain-Shams University Organization of African Unity Street Cairo Abassia 11566 Egypt Research Center for Chinese Herbal Medicine College of Human Ecology Chang Gung University of Science and Technology Taoyuan 33303 Taiwan Graduate Institute of Health Industry Technology College of Human Ecology Chang Gung University of Science and Technology Taoyuan 33303 Taiwan Department of Anaesthesiology Chang Gung Memorial Hospital Taoyuan 33305 Taiwan Graduate Institute of Natural Products College of Medicine Chang Gung University Taoyuan 33302 Taiwan Department of Chemical Engineering Ming Chi University of Technology New Taipei City 24301 Taiwan National Museum of Marine Biology & Aquarium Pingtung 94450 Taiwan Department of Marine Biotechnology and Resources National Sun Yat-sen University Kaohsiung 80424 Taiwan Traditional Herbal Medicine Research Center Taipei Medical University Hospital Taipei 11031 Taiwan

Recent advancements in the discovery of natural products have made significant progress in rapidly identifying known compounds from complex extracts using advanced spectroscopic technologies. Among these methods, one powerful approach called MS/MS molecular networking (MN) organizes chemical similarity based on MS/MS fragmentation data, serving as a valuable complement to traditional dereplication strategies. In our ongoing research focused on lead discovery from a sustainable medicinal source, an aquaculture soft coral Sarcophyton trocheliophorum, the MN profiling approach revealed the chemical diversity of biscembranoid analogs. By analyzing the MN pattern, biscembranoid derivatives with unique MS/MS fragments were prioritized for further investigation. Two novel biscembranoids, sarcotroxide A (1) and B (2), were successfully obtained through targeted isolation. These novel biscembranes contained an unprecedented 3-hydroperoxy-3-methylcyclohex-1-ene moiety compared with all biscembranoid analogs. The chemical structures of the two new biscembranoids were confirmed through spectroscopic analyses and in silico calculations. Their anti-inflammatory properties were assessed against the generation of superoxide anion (O₂^•−) and elastase release in activated human neutrophils. Sarcotroxides A (1) and B (2) exhibited anti-inflammatory activity by inhibiting O₂^•− accumulation (with 43.67% and 61.62% inhibition) and elastase release (with 38.21% and 61.62% inhibition) at 10 µM. These findings validated the potential of these biscembranoids from the cultured S. trocheliophorum as promising candidates for the future development of anti-inflammatory agents specifically designed to target neutrophils. © 2023 The Authors

关键词： Anti-inflammation Aquaculture soft coral Biscembranoid Neutrophil Sarcophyton trocheliophorum

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The association of combinations of social factors and SARs-CoV-2 infection: A retrospective population-based cohort study in Ontario, 2020–2021

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Dialogues in Health 2024年 5卷 100197-100197页

作者： Persaud, Sydney Fitzgerald, Michael Hawken, Steven Tanuseputro, Peter Boucher, Lisa Petrcich, William Wellman, Martin Webber, Colleen Shoemaker, Esther Ducharme, Robin Dahrouge, Simone Myran, Daniel Bayoumi, Ahmed M. Wanigaratne, Susitha Bloch, Gary Ponka, David Smith, Brendan T. Lofters, Aisha Zygmunt, Austin MacLeod, Krystal Kehoe Turcotte, Luke A. Sander, Beate Howard, Michelle Funnell, Sarah Rayner, Jennifer Kitagawa, Kurtis Ibrahim, Sureya Kendall, Claire E. Bruyère Health Research Institute Ottawa ON Canada ICES Ottawa ON Canada Ottawa Hospital Research Institute Ottawa ON Canada School of Epidemiology and Public Health University of Ottawa Ottawa ON Canada Department of Family Medicine University of Ottawa Ottawa ON Canada Institute of Health Policy Management and Evaluation University of Toronto Toronto ON Canada Li Ka Shing Knowledge Institute St. Michael's Hospital Toronto ON Canada Division of General Internal Medicine St. Michael's Hospital Toronto ON Canada Department of Medicine University of Toronto Toronto ON Canada Institute of Medical Sciences University of Toronto Toronto ON Canada Edwin S.H. Leong Center for Healthy Children University of Toronto Toronto ON Canada Dalla Lana School of Public Health University of Toronto Toronto ON Canada Unity Health Toronto ON Canada Public Health Ontario Toronto ON Canada Department of Family and Community Medicine University of Toronto Toronto ON Canada Department of Health Sciences Brock University St. Catharines ON Canada Toronto General Hospital Research Institute University Health Network Toronto ON Canada Department of Family Medicine McMaster University Hamilton ON Canada Faculty of Health Sciences Queen's University Kingston ON Canada Department of Family Medicine Queen's University Kingston ON Canada Alliance for Healthier Communities Toronto ON Canada Centre for Community Learning & Development Toronto M5A 2B3 ON Canada

Objective: The COVID-19 pandemic highlighted and exacerbated health inequities worldwide. While several studies have examined the impact of individual social factors on COVID infection, our objective was to examine how interactions of social factors were associated with the risk of testing positive for SARS-CoV-2 during the first two years of the pandemic. Study design and setting: We conducted an observational cohort study using linked health administrative data for Ontarians tested for SARS-CoV-2 between January 1st, 2020, and December 31st, 2021. We constructed multivariable models to examine the association between SARS-CoV-2 positivity and key variables including immigration status (immigrants vs. other Ontarians), and neighbourhood variables for household size, income, essential worker status, and visible minority status. We report main and interaction effects using odds ratios and predicted probabilities, with age and sex controlled in all models. Results: Of 6,575,523 Ontarians in the cohort, 88.5 % tested negative, and 11.5 % tested positive for SARS-CoV-2. In all models, immigrants and those living in neighbourhoods with large average household sizes had greater odds of testing positive for SARS-CoV-2. The strength of these associations increased with increasing levels of neighbourhood marginalization for income, essential worker proportion and visible minority proportion. We observed little change in the probability of testing positive across neighbourhood income quintiles among other Ontarians who live in neighbourhoods with smaller households, but a large change in probability among other Ontarians who live in neighbourhoods with larger households. Conclusion: Our study found that SARS-CoV-2 positivity was greater among people with certain combinations of social factors, but in all cases the probability of testing positive was consistently greater for immigrants than for other Ontarians. Examining interactions of social factors can provide a more nuance

关键词： COVID-19 positivity Health equity SARS-CoV-2 Social determinants of health

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Notable intestinal dysbiosis orchestrated by Escherichia/Shigella, decreased levels of SCFTA (short chain fatty acids) and alterations in metabolic pathways characterize patients with alcohol-decompensated cirrhosis in western Mexico

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Annals of Hepatology 2022年 27卷 100812-100812页

作者： Baltazar-Díaz, TA González-Hernández, LA Aldana-Ledesma, JM Peña-Rodríguez, M Vega-Magaña, AN Zepeda-Morales, ASM López-Roa, RI Toro-Arreola, S Del Bueno-Topete, MR Chronic-Degenerative Diseases Research Institute. University Center for Health Sciences. Guadalajara University. Guadalajara. Jalisco VIH Unity. Hospital Civil Fray Antonio Alcalde. Guadalajara Jalisco Gastroenterology Service. Hospital Civil Fray Antonio Alcalde. Guadalajara. Jalisco Diagnostic Laboratory for Emerging and Reemerging Diseases. University Center for Health Sciences. Guadalajara University. Guadalajara. Jalisco. México Research Institute in Biomedical Sciences. Guadalajara University. Guadalajara. Jalisco. México Pharmaceutical Research and Development Laboratory. University Center of Exact Sciences and Engineering. Guadalajara University. Guadalajara. Jalisco. México

Introduction and ObjectivesTo evaluate the composition and functions of the intestinal microbiota in patients with alcohol-decompensated cirrhosis. Materials and methodsFecal samples of eighteen patients and eighteen healthy controls (HC) were obtained. Microbial composition was characterized by 16S rRNA amplicon sequencing, SCFAs quantification was performed by gas chromatography (GC), metagenomic predictive profiles were analyzed by PICRUSt2. ResultsGut microbiota in the cirrhosis group revealed a significant increase in the pathogenic genera Escherichia/ Shigella and Prevotella, a decrease in beneficial bacteria, such as Blautia, Faecalibacterium, plus a decreased α-diversity ( p<0.001) compared to HC. Fecal SCFAs concentrations were significantly reduced in the cirrhosis group ( p<0.001). PICRUSt2 analysis indicated a decrease in acetyl-CoA fermentation to butyrate, as well as an increase in pathways related to antibiotics resistance and aromatic amino acid biosynthesis. DiscussionThe gut microbiota dominated by the Escherichia/ Shigella general correlates with low SCFA concentrations and an increase in metabolic pathways related to pathogenicity and the production of substances associated with endotoxemia. ConclusionsGut microbiota of these patients possesses a pathogenic/inflammatory environment. Therefore, future strategies to balance intestinal dysbiosis should be implemented. These findings are described for the first time in the population of western Mexico. FundingThe resources used in this study were from the hospital without any additional financing Declaration of interestThe authors declare no potential conflicts of interest.

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CD4+ tissue-resident memory Th17 cells are a major source of IL-17A in Spondyloarthritis synovial tissue

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Annals of the rheumatic diseases 2025年

作者： Feng Liu Hui Shi Jason D Turner Rachel Anscombe Jiaqi Li Takuya Sekine Ariane Hammitzsch Devika Agarwal Christopher Mahony Jiewei Chen Ben Kendrick Dajiang Du Qiang Tong Lihua Duan Kyla Dooley Hai Fang Ilya Korsunsky Roopa Madhu Adam P Cribbs Matthias Friedrich Brian D Marsden Yi-Ling Chen Graham Ogg Anna Adams Warner Chen Steven Leonardo Fiona E McCann Christopher D Buckley Terence Rooney Thomas Freeman Holm H Uhlig Calliope Dendrou Adam Croft Andrew Filer Paul Bowness Liye Chen Nuffield Department of Orthopaedics Rheumatology and Musculoskeletal Sciences University of Oxford Oxford United Kingdom. Rheumatology Research Group Institute of Inflammation and Ageing University of Birmingham Birmingham United Kingdom. Kennedy Institute of Rheumatology Old Road Campus University of Oxford Oxford United Kingdom. Center for Infectious Medicine Department of Medicine Huddinge Karolinska Institutet Stockholm Sweden. TUM University Hospital Klinikum Rechts der Isar Department of Nephrology Klinikum Rechts der Isar School of Medicine and Health Technical University of Munich Munich Germany. Rheumatology Research Group Institute of Inflammation and Ageing University of Birmingham Birmingham United Kingdom NIHR Birmingham Biomedical Research Center and Clinical Research Facility University of Birmingham Queen Elizabeth Hospital Birmingham United Kingdom. Shanghai Sixth People's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine. Nuffield Orthopaedic Centre Oxford University Hospitals Trust Oxford United Kingdom Nuffield Department of Orthopaedics Rheumatology and Musculo-skeletal Science University of Oxford Oxford United Kingdom. Shanghai Sixth People's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine Division of Rheumatology and Clinical Immunology Department of Medicine Queen Mary Hospital Hong Kong China. Department of Rheumatology and Clinical Immunology Jiangxi Provincial People's Hospital The First Affiliated Hospital of Nanchang Medical College Nanchang China. Translational Gastroenterology Unit University of Oxford Oxford United Kingdom. Shanghai Institute of Hematology State Key Laboratory of Medical Genomics National Research Center for Translational Medicine at Shanghai Ruijin Hospital Shanghai Jiao Tong University School of Medicine Shanghai 200025 China. Brigham and Women's Hospital Boston MA USA Harvard Medical School Boston MA USA Broad Institute of Massachusetts Institute of Techno

OBJECTIVES:Interleukin (IL)-17A is a key driver of spondyloarthritis (SpA) joint pathology. We aimed to identify its cellular source in synovial tissue from patients with 2 forms of SpA, namely axial SpA (AxSpA) and psoriatic arthritis (PsA). METHODS:Synovial tissue from patients with SpA was profiled using single-cell RNA sequencing (scRNA-seq; AxSpA, n = 5; PsA, n = 6) or spatial RNA profiling (PsA, n = 4). CellPhoneDB was used to infer cell-cell communication. Tissue-resident memory Th17 (TRM17)-like cells were generated in vitro using blood memory CD4+ T cells from SpA patients. An epigenetic inhibitor library, siRNA, and clustered regularly interspaced short palindromic repeats (CRISPR) were used to identify epigenetic regulator(s) for TRM17. RESULTS:scRNA-seq showed that CD4+CXCR6+ TRM17 cells are the predominant spontaneous IL17A producers in SpA synovium. Cell-cell communication and single-cell spatial analysis support the interaction between TRM17 and CLEC10A+ dendritic cells, which were activated in SpA. Both sublining and lining fibroblasts in SpA synovium showed evidence of interleukin (IL)-17A activation. In vitro-generated CD4+ TRM17-like cells phenocopied joint tissue TRM17, producing IL-17A/F upon T cell-receptor (TCR) stimulation, which was further enhanced by cytokines. Perturbation of BRD1 inhibited the generation of TRM17-like cells. CONCLUSIONS:CD4+ TRM17 cells are the predominant source of IL-17A in SpA synovial tissue. TCR stimulation is essential for the secretion of IL-17A by CD4+TRM17-like cells. The epigenetic regulator bromodomain-containing protein 1 (BRD1) contributes to the generation of CD4+TRM17. Depleting CD4+TRM17 cells in SpA is thus a therapeutic strategy with potential to induce long-term remission.

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Synthesis and characterization of ZSM-5/MCM-41 composite molecular sieves

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Ranliao Huaxue Xuebao/Journal of Fuel Chemistry and Technology 2013年第10期41卷 1268-1273页

作者： Zhang, Jun-Tao Hao, Na-Na Wang, Ni Research Center of Petroleum Processing and Petrochemicals Xi'an Shiyou University Xi'an 710065 China Shanghai New-Unity Energy Technology Co. LTD Shanghai 201210 China Hanzhong Natural Gas Investment Development Co. LTD Hanzhong 723000 China

ZSM-5/MCM-41 composite molecular sieves with multiple micro-mesoporous structure were hydrothermally synthesized via self-assembly by using the alkali-treated ZSM-5 seriflux as the source of silica and aluminum. The as-synthesized molecular sieves were characterized by XRD, N2 adsorption, HRTEM, Py-IR, and hydrothermal treatment methods. The results showed that the crystallinity of the as-synthesized molecular sieves is dependent on the intensity of the alkali-treatment;the suitable alkali treatment conditions for ZSM-5 are 80°C for 1 h with a NaOH concentration of 1 mol/L. The composite molecular sieves obtained exhibit a typical MCM-41 structure, with a hierarchical micro-mesoporous structure and large specific surface area. Compared with MCM-41, the ZSM-5/MCM-41 composite molecular sieves show higher quantity of Bronsted acid sites (especially strong ones) and higher hydrothermal stability.

关键词： Hydrothermal synthesis

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