55,978 research outputs found

    Biocontrol as a key component to manage brown rot on cherry

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    Brown rot, caused by Monilinia spp., is one of the most important diseases in stone fruits worldwide. Brown rot can cause blossom wilts and fruit rots in the orchard as well as latent infections of fruit, leading to post-harvest fruit decaying. Current control methods rely on scheduled spraying of fungicides. However, the continuing pressure to reduce fungicide use has seen an increase in research into alternative management methods, such as biological control. NIAB EMR recently identified two microbes that significantly reduced sporulation of Monilinia laxa under laboratory conditions. These two isolates were a bacterial species Bacillus subtilis (B91) and yeast-like fungus Aureobasidium pullulans (Y126) and are currently being formulated into commercial products. We are investigating how to optimise the use of these two potential biocontrol products in practice, in terms of suppressing Monilinia sporulation on overwintered mummies and preventing infection of blossoms and fruits. When applied to mummified fruits in winter Y126鈥檚 population was stable through the winter but at a low concentration. The B91 survived a little longer with the population reaching that of the control group by week 4. Neither Biological control (BCA) treatments had an affected the population of M. laxa when compared to the control treatment of sterile distilled water. The interaction time between the BCAs and M. laxa showed the longer the interaction time the lower the spore count of M. laxa. Another study was performed looking into the ability of our BCAs to colonise and survive on blossoms. B91 did not survive well on blossoms but could survive on fruits. However, its antagonistic compounds need to be in relatively high concentration to be effective against M. laxa. Therefore, it is best used as a fungicide, ensuring the antagonistic compounds are at a high concentration when applied in the field. Y126 can persist throughout the season and was marginally, though not statistically significantly, more effective at long term reduction in M. laxa. This could be because Y126 works through competition, therefore the interaction time with the pathogen could be important for efficacy and something worth investigating further. The difference between the BCAs highlights the need to understand each BCA鈥檚 ecology to ensure maximum efficacy. In a latent infection experiment, we inoculated trees with M. laxa and then treated them with the two biocontrol isolates two weeks before harvest. Post-harvest disease development was assessed after four days of storage in 2019 and two weeks in 2020. There was a significant reduction in rot incidence (p < 0.001) of 29% (Y126) and 27% (B91) in 2019 and 62 % (Y126) and 80 % (B91) in 2020 when the harvested fruit was stored at cold store levels. With new products to be introduced into the environment, it's important to understand the effects they may have on the plant's microbiome. Using next-generation sequencing techniques, we looked at the impact B91 and Y126 has on the blossom and cherry microbiomes. There was a treatment effect in both the bacterial and fungal communities on the blossom and ripe cherry. But the biggest variability was between blocks (Geographical effect) and between the years in which we experimented (p < 0.0001). This research will assist in the development of management strategies, especially spray timings for brown rot on stone fruit, integrating BCAs with other management practices

    Mapping Super-Relaxed States of Myosin Heads in Sarcomeres using Oblique Angle Fluorescent Microscopy

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    We have utilised modern methods of super-resolution fluorescent microscopy to spatially map fluorescently labelled ATP molecules in relaxed rabbit psoas skeletal muscles. For our imaging process, we have labelled ATP molecules with Rhodamine and Z-lines with Alexa488. Data from imaging these fluorophores have been collected using oblique angle fluorescent microscopy and further analysed to map super relaxed states (SRX) of myosin heads on the thick filament. Our experiments have concluded that most SRX of myosin heads were found in the C-zone of the thick filament, while other zones of thick filament had smaller populations of SRX. Further introduction of mavacamten (MAVA) to our imaging system has revealed an increase in SRX in both D and P zones, while the C zone population of SRX had remained constant. Further experiments must be conducted to establish a clear pattern and further proof our findings

    Bioinformatic characterization of a triacylglycerol lipase produced by Aspergillus flavus isolated from the decaying seed of Cucumeropsis mannii

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    Lipases are enzymes of industrial importance responsible for the hydrolysis of ester bonds of triglycerides. A lipolytic fungus was isolated and subsequently identified based on the ITS sequence analysis as putative Aspergillus flavus with accession number LC424503. The gene coding for extracellular triacylglycerol lipase was isolated from Aspergillus flavus species, sequenced, and characterised using bioinformatics tools. An open reading frame of 420 amino acid sequence was obtained and designated as Aspergillus flavus lipase (AFL) sequence. Alignment of the amino acid sequence with other lipases revealed the presence GHSLG sequence which is the lipase consensus sequence Gly-X1-Ser-X2-Gly indicating that it a classical lipase. A catalytic active site lid domain composed of TYITDTIIDLS amino acids sequence was also revealed. This lid protects the active site, control the catalytic activity and substrate selectivity in lipases. The 3-Dimensional structural model shared 34.08% sequence identity with a lipase from Yarrowia lipolytica covering 272 amino acid residues of the template model. A search of the lipase engineering database using AFL sequence revealed that it belongs to the class GX-lipase, superfamily abH23 and homologous family abH23.02, molecular weight and isoelectric point values of 46.95鈥塊Da and 5.7, respectively. N-glycosylation sites were predicted at residues 164, 236 and 333, with potentials of 0.7250, 0.7037 and 0.7048, respectively. O-glycosylation sites were predicted at residues 355, 358, 360 and 366. A signal sequence of 37 amino acids was revealed at the N-terminal of the polypeptide. This is a short peptide sequence that marks a protein for transport across the cell membrane and indicates that AFL is an extracellular lipase. The findings on the structural and molecular properties of Aspergillus flavus lipase in this work will be crucial in future studies aiming at engineering the enzyme for biotechnology applications

    Uso de las histonas circulantes y sus modificaciones post-traduccionales como biomarcadores en sepsis y shock s茅ptico

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    La sepsis es una afecci贸n potencialmente mortal causada por una respuesta anormal del hu茅sped a una infecci贸n, produciendo respuestas fisiol贸gicas alteradas que da帽an los propios tejidos del paciente y pueden provocar disfunci贸n org谩nica e incluso la muerte. Asimismo, algunos pacientes s茅pticos progresan a shock s茅ptico, caracterizado por alteraciones circulatorias, celulares y metab贸licas sustanciales que aumentan el riesgo de mortalidad. A pesar de que la sepsis se caracteriza por un mal funcionamiento del sistema inmunol贸gico, lo que a su vez conduce a una respuesta inmune alterada e inmunosupresi贸n, la alta complejidad de la fisiopatolog铆a de la sepsis requiere una mayor investigaci贸n para comprender las respuestas inmunes que ocurren durante la sepsis. Asimismo, las histonas extracelulares circulantes han ganado relevancia como mediadores citot贸xicos en la sepsis, ya que act煤an como patrones moleculares asociados a da帽o, que inducen estr茅s oxidativo y activan el inflamasoma NLRP3. Estos mecanismos median la activaci贸n de la piroptosis, un mecanismo de muerte celular programada que produce inflamaci贸n mediante la expresi贸n de IL-18, IL-1尾 and IL-1伪. Sin embargo, a pesar de la evidencia de activaci贸n del inflamasoma en las c茅lulas inmunes durante la sepsis, se desconoce si las histonas extracelulares son capaces de activar los inflamasomas endoteliales y sus consecuencias. En este trabajo destacamos el papel previamente desconocido de las histonas extracelulares, mediando la activaci贸n del inflamasoma NLRP3 y la piroptosis en las c茅lulas endoteliales, contribuyendo a la disfunci贸n endotelial y la desregulaci贸n de la respuesta inmune mediada por el endotelio. Asimismo, tambi茅n demostramos c贸mo la acetilaci贸n de histonas disminuye la activaci贸n de la piroptosis. Adem谩s, demostramos que la piroptosis se produce en pacientes con shock s茅ptico y los niveles de histonas circulantes se correlacionan con la expresi贸n de citoquinas proinflamatorias y citoquinas piropt贸ticas, la liberaci贸n de factores de adhesi贸n endotelial y la gravedad de la enfermedad. Proponemos la piroptosis mediada por histonas como un nuevo objetivo para desarrollar intervenciones cl铆nicas. De manera similar, hemos analizado las respuestas inmunorelacionadas que ocurren durante las primeras etapas de la sepsis con el objetivo de proporcionar nuevos datos comparando las cantidades de citoquinas, inmunomoduladores y otros mediadores endoteliales en pacientes cr铆ticamente enfermos no s茅pticos, s茅pticos y de shock s茅ptico. Nuestro enfoque ayudar谩 a caracterizar r谩pidamente las respuestas inmunes alteradas en pacientes s茅pticos y de shock s茅ptico ingresados en la Unidad de Cuidados Intensivos. Finalmente analizamos el papel de la metilaci贸n del ADN en el control del sistema inmune s茅ptico. Nuestros resultados demostraron el papel central de la metilaci贸n del ADN modulando la respuesta molecular en los pacientes de shock s茅ptico y contribuyendo a la inmunosupresi贸n, a trav茅s de la alteraci贸n de los patrones de metilaci贸n de los promotores de IL-10 y TREM-2.Sepsis is a life-threatening condition caused by an abnormal host response to an infection that produce altered physiological responses which damages own tissues of the patient and can result in organ dysfunction and in some cases death. Likewise, a subset of septic patients progresses to septic shock, characterized by substantial circulatory, cellular and metabolic abnormalities, which substantially increase the risk of mortality. Sepsis is characterized by a malfunction of the immune system and it can lead to an altered immune response and immunosuppression. Moreover, the high complexity of the pathophysiology of sepsis requires of further investigation to characterize the immune responses in sepsis and septic shock. Likewise, circulating extracellular histones have gained relevance as cytotoxic mediators in sepsis pathophysiology, since they act as damage-associated molecular patterns, which induce oxidative stress and activate NLRP3 inflammasome. Subsequently, inflammasome mediates pyroptosis activation, a programmed cell death mechanism that produces inflammation through the release of IL-18, IL-1尾 and IL-1伪. However, despite inflammasome activation may occur in immune cells during sepsis, it is unknown if this process also takes place in endothelial cells and particularly whether extracellular histones are capable of activating endothelial inflammasomes and their consequences. In this work we highlight a previously unknown role for extracellular histones, that mediates the activation of NLRP3 inflammasome and pyroptosis in endothelial cells by contributing to endothelial dysfunction and the dysregulation of the immune response mediated by endothelium. Likewise, we demonstrated how histone acetylation decreases pyroptosis activation. Furthermore, we show how pyroptosis occurs in septic shock patients and how circulating histone levels correlate with the expression of pro-inflammatory and pyroptotic cytokines, the release of endothelial adhesion factors and septic shock severity. We propose histone-mediated pyroptosis as a new target to develop clinical interventions. Similarly, we have analyzed the immune-related responses occurring during the early stages of sepsis with the aim of providing new data by comparing the amounts of cytokines, immune modulators and other endothelial mediators in critically-ill non-septic patients, septic and septic shock patients. Our approach will help to rapidly characterize the altered immune responses in septic and septic shock patients admitted in the Intensive Care Unit. Finally, we also analyzed the role of DNA methylation in the control of septic immune system. Our results demonstrated the central role of DNA methylation modulating the molecular response in septic shock patients and contributing to immunosuppression, through the alteration of DNA methylation patterns of IL-10 and TREM2 promoters

    Conservation agriculture affects grain and nutrient yields of Maize (Zea Mays L.) and can impact food and nutrition security in Sub-Saharan Africa

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    Maize is a major staple and plays an essential role in food and nutrition security in Sub-Saharan Africa (SSA). Conservation agriculture (CA), a climate-smart agriculture practise based on minimum soil disturbance, crop residue retention, and crop diversification, has been widely advocated but without extensive research on the impact it may have on maize nutrient composition, and food and nutrition security. This study assessed the grain yield, macro- and micronutrient mineral content, and nutrient yield of eight maize varieties grown in Malawi, and how these are affected by CA practises over two seasons. The minerals were analysed by inductively coupled plasma (ICP) coupled to optical emission spectroscopy (OES) and to mass spectroscopy (MS). Grain yield and Se content differed among the varieties, while C, N, Fe, K, Mg, Mn, P, and Zn were similar. The local variety Kanjerenjere showed lowest grain and nutrient yields. The open-pollinated varieties (OPVs) concentrated more minerals than the F1 hybrids, but the latter showed higher yields for both grain and nutrients. Typical consumption of the eight maize varieties could fully meet the protein and Mg dietary reference intake (DRIs) of Malawian children (1鈥3 years), as well as Mg and Mn needs of adult women (19鈥50 years), but their contribution to dietary requirements was low for Fe (39鈥41%) and K (13鈥21%). The trials showed that CA increased grain yield (1.2- to 1.8-fold) and Se content (1.1- to 1.7-fold), but that it had no effect on C, K, Mg, P, and Zn, and that N (1.1- to 1.2-fold), Mn (1.1- to 1.8-fold), and Fe (1.3- to 3.4-fold) were reduced. The high increase in grain yield under CA treatments resulted in increased yields of protein and Se, no effect on the yields of K, Mg, Mn, P, Zn, and reduced Fe yield. Conservation agriculture could contribute in reducing the risk of Se deficiency in Malawian women and children but exacerbates the risk of Fe deficiency. A combination of strategies will be needed to mitigate some of the foreseen effects of climate change on agriculture, and food and nutrition security, and improve nutrient intake

    The Caribbean Syzygy: a study of the novels of Edgar Mittelholzer and Wilson Harris

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    The problem of racial inheritance - the "search for identity" - is a recurring theme in the criticism of Caribbean literature. It is a pre-occupation with Caribbean writers, affecting both subject matter and literary quality, as FM. Birbalsingh, for example, has shown with reference to the novels of John Hearne and E,R. Braithwaite (Caribbean quarterly Vols. 14, December 1968 and 16, March 1970). This study of the work of Edgar Mittelholzer and Wilson Harris will attempt to show that there are important areas still to be explored relating Caribbean literature to its complex racial and cultural background. Both Mittelholzer and Harris deserve close, critical study in their own right; but a parallel examination reveals similarities and differences which bring into sharper focus wider concerns of Caribbean literature. The two important directions of West Indian writing are more clearly seen: the one, pioneered by Mittelholzer, in which the writer looks outward towards a "parent" culture, and the other looking inward, seeking in its own, complex inheritance the raw material for new and original growth. Mittelholzer and Harris are both Guyanese of mixed racial stock, both deeply concerned with the psychological effects of this mixture, and both writers have a profound awareness of the Guyanese historical and cultural heritage. They also share a deep feeling for the Guyenese landscape which appears in their work as a brooding presence affecting radically -the lives of those who live within i-t. Mittelholzer's attitude to his mixed racial and cultural origins, however, produces in his work a schizophrenic Imbalance while Harris, by accepting racial and cultural complexity as a starting-point, initiates a uniquely creative and experimental art. Mittelholzer, in his approach to history, human character eM landscape, remains a vi "coastal" writer never really concerned (as Harris is) with. the deeper significance of the "Interior" and all that this implies, both in a geographical and psychological sense. The fact that Mittelbolzer's work reflects a psychological imbalance induced by a pre-occupation with racial identity has been demonstrated by Denis Williams in the 1968 Mittelholzer Lectures, and by Joyce Sparer in a series of articles in the Guyana Graphic. Mittelholzer's awareness of this imbalance, however, and his attempt to come to terms with it in his art remain to be examined and documented, as does Harris's attempt to create am "associative" art aimed at healing the breach in the individual consciousness of Caribbean Man. The aim of this study is to demonstrate that Mitteholzer and. Harris, although antithetical in impact and style (each representing an approach to fiction directly opposed to the other) are, in fact, the opposite elements of a dichotomy. Their work illustrates the negative and positive aspects of the racial and cultural schizophrenia of the Caribbean, for both writers in their different ways are preoccupied with (and therefore have embodied in their work) the juxtaposition and, contrasting of apparently irreconcilable emotional and intellectual qualities - the Caribbean Syzygy

    Gendered spaces in contemporary Irish poetry

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    The thrust of this thesis is summarized by the following questions: How does contemporary Irish poetry migrate from traditional conceptions of identity drawn on by the cultural nationalism of the Irish Literary Revival, and what effects does this have on understanding gendered and national identity formation? Chapters are on the following: Seamus Heaney, Tom Paulin, Paul Muldoon, MedbhMcGuckian, Eavan Boland and Sara Berkeley. These poets are chosen for discussion since their work most effectively engages with the relationship between woman and nation, the representation of gendered national identity, and the importance of feminist and post-colonial theorization. Focusing on poetry worth and South of the border from the last fifteen years, the thesis asks how a younger generation of poets provide a response to nationality which is significantly different from their predecessors. The thesis is composed of three parts: the first understand how the male poets depart from conventional conceptions of the nation with reference to post-colonial theorization; the second explores how feminist theorization informs readings of how the female poets respond to the nation; the final part investigates migration in the poetry and problematizes this in terms of post-nationalism. Discussing the issue of deterritorialization in Irish poetry, the thesis notice how as the poets attempt to take flight from the mythologies of nationhood, they undermine the monoliths of gendered and national identity inscribed within Irish political discourse, which is typified at a representative level by the figure of Mother Ireland or Cathleen Ni Houlihan. Investigating the ways in which gender and nation, and the body and space are reinscribed by the poets, the thesis argues that their poetry challenges authentic conceptions of Irish identity and the nation-state, so as to loosen the legacy of a colonial and nationalist inheritance

    Towards the development of care management in community care for elderly people in Korea

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    This study is concerned with the feasibility of several forms of care management in the development of community care for elderly people in Korea. Chapter one introduces the background of community care in Korea in the light of demographic, socio-economic, and political realities. This chapter reviews the changing Korean society as a barometer to understand the scope, size, and speed of social needs, especially community care for elderly people, over the last few decades. Chapter two explores various definitions, concepts, and theories of community, community care, and care management by building upon trends previously established in the research. This helps to identify the different models of care management and the pre-conditions necessary for the application of different models in Korea. Chapter three explores what factors have affected the development of community care, and what community dare has achieved for elderly people in the UK. Especially, care management in community care for elderly people in the UK is examined in detail. Chapter four details the findings of field research on community care for elderly people in Korea. This covers the needs of elderly people and their carers, and the social worker's tasks and available resources. The potential for the use of care management based on the findings of field research is assessed. Chapter five investigates whether the UK models of care management are suitable for Korean society, which interventions are useful for developing care management, and the strategies, and principles involved

    Investigating the potential role for RBMY in cancer

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    Introduction: Head and neck squamous cell carcinoma (HNSCC), is a major healthcare concern with a high male prevalence. We hypothesise that the testis specific mRNA splicing regulator, Y-linked RBMY gene, is aberrantly expressed in HNSCC in part promoting HNSCC through ZFY-short splicing. RBMY has been shown to enhance tumour development in male hepatocellular carcinoma (human tissue specimens and transgenic-mouse models) whilst ZFY-short is predicted to have anti-apoptotic properties and the deletion of RBMY locus on Y-chromosome resulted in lowered ZFY-short expression. Thus, we hypothesize that ZFY-short is generated by RBMY and exerts its anti-apoptotic effects to promote male HNSCC. Methods: Due to the coronavirus lockdown, bench work was restricted to 6 months, therefore, I conducted an extended analysis of RBMY expression in human cancer, including a computational analysis of RBMY gene expression with data from the cBioPortal database. In my bench-work, I attempted to establish GFP- RBMY expressing cell lines and conducted fluorescence microscopy, RT- PCR and qPCR to analyse RBMY expression in HNSCC cell lines and its impact on ZFY-short expression. Results: RBMY is expressed in several cancers, with no driver mutations. RBMY has nuclear localisation and is expressed in 93-UV-147T and UM-SCC-104 cell lines (both HPV16-positive HNSCC cell lines), with increased ZFY-short expression observed in UM- SCC-104. Discussion: Despite RBMY having been shown to be an oncogene in male liver cancer, our analysis of cBioPortal data suggests this activity may be restricted to the small minority of tumours of different cancer types that express RBMY. The paralleled expression of RBMY and ZFY-short in our cell lines indicate an association. UMSCC104 cell line originates from a highly an aggressive and recurrent tumour, RBMY is associated with tumour stemness, thus it is possible that via ZFY-short, RBMY could have promoted the aggressive phenotype in this, and in other HNSCCs
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