Academic literature on the topic '90-90-90 Target'

The prevalence of human immunodeficiency virus (HIV) infection is low but it is steadily increasing in Turkey. In the current study, we aimed to assess the status of HIV infection management with the proposed 90–90–90 targets in a large HIV cohort in Istanbul, Turkey. The cohort included 2382 patients (2082 male, 300 female, mean age was 36.3 ± 11.3 years). Mean CD4 cell count was 399 cells/mm3 and HIV-RNA level was 576,235 copies/ml. According to the modeling by the Modeling tool of European Center for Diseases Control Software, 72 and 74% of all HIV patients had been diagnosed in 2016 and 2017, respectively (the first target). Among 2382 patients, 2191 (92%) were on antiretroviral therapy (the second target). The third target of virally suppressing those on treatment was achieved among 70.2% of the patients. The current study suggests that both the fraction of those living with undiagnosed HIV and the proportion of those on treatment who are virally suppressed should be targeted to sustain optimal HIV care. Efforts should continue to surpass the targets of 90–90–90.

Despite progress in the prevention and treatment of HIV, persistent issues concerning the evaluation of continuum in care from the serological diagnosis to virologic success remain.
 Considering the UNAIDS target 90-90-90 for 2020 for treatment and viral suppression of people living with HIV (PLVH), our purpose was to verify if, starting from diagnosis, the viral suppression rate of our cohort of new PLWH satisfied the above targets.
 The aim of this retrospective study was to compare 2005-2017 data collected at the Perugia Infectious Diseases Clinic with the 2020 UNAIDS 90 targets and to identify risk factors that could be associated with failure to reach these targets.
 Methods: We included all patients aged ≥15 undergoing HIV test at our clinic between January 2005 and December 2017. We evaluated the unclaimed tests, linkage to care, retention in ART and the viral suppression at 1 and 2 years from starting ART. Data were analyzed between Italians and foreigners.
 Results: We observed 592 new diagnoses for HIV infection: 61.4% on Italian-natives, 38.5% on foreigners. Considering the continuum of care from diagnosis, 88 (15%) PLWHIV were lost to engagement in care: 55 (9.2%) patients didn’t withdraw the test and 33 (5.5%) didn’t link to care.
 An antiretroviral treatment was started only on 78.8% of the new diagnoses (467/592) and a viral suppression was obtained at 2 years on 82% of PLWH who had started ART (383/467) namely only 64.7% of the new diagnoses instead of the hoped-for 81% of the UNAIDS target. We found no significant differences between Italians and foreigners
 Conclusions
 UNAIDS goal was very far to be reached. The main challenges were unreturned tests as well as the retention in ART. Rapid tests for a test-treat strategy and frequent phone communications in the first ART years could facilitate UNAIDS target achievement.

Background: In 2016, Khayelitsha reported almost 89% viral load (VL) suppression but less than 56% completion on routine quarterly reports, casting doubt on the validity of reported suppression. Objectives: To assess the validity of reported VL suppression as a measure of progress towards the 90-90-90 target and identify barriers to routine VL completion. Methods: A retrospective cohort study including all patients on antiretroviral treatment (ART) in Khayelitsha with a routine VL expected between 1 July 2015 and 30 June 2016 was conducted. ART programme and laboratory data were obtained and a sample of 1 035 patient folders were reviewed. Suppression was calculated using laboratory data and compared to reported suppression. A VL cascade from “expected” to “done”, “filed”, “noted” and “captured” was constructed to reflect the steps a VL must complete to be included in reported suppression and successful progression to each step was estimated. Logistic regression models were used to calculate adjusted odds ratios (aOR) with 95% confidence intervals (95%CI) for completion among different patient groups. Results: Using laboratory data, VL suppression was estimated to be 82%, 87%, 89% and 91% at the 50, 200, 400 and 1 000 copies/mL thresholds respectively, but reported suppression would have been 80%, 86%, 88% and 89% at those thresholds. Of 22 991 patients with a routine VL due in the study period, 84% were done, 79% filed, 76% noted, and 55% captured. Routine VL were more likely to be done among children< 15 years old (aOR 1.89, 95%CI 1.45–2.48) and pregnant women (aOR 1.90, 95%CI 1.28–2.81) compared to adult men, adjusted for facility. Conclusion: Despite low reported completion, actual completion was high and reported suppression was similar to suppression calculated using laboratory data, thus providing an accurate measure of progress towards the 90-90-90 target. More work is needed to reach the 16% of patients missed by routine testing. Most of the VL done were available to the clinician at the next assessment, and further research is needed to assess how effectively these VL results are used in clinical decision-making.

許諾条件により本文は公開していません : 本文は「The Journal of Neuroscience. January 1. 1999, 79(1) : pp.109-118」に掲載<br>本文データは平成22年度国立国会図書館の学位論文(博士)のデジタル化実施により作成された画像ファイルを基にpdf変換したものである<br>Kyoto University (京都大学)<br>0048<br>新制・課程博士<br>博士(医学)<br>甲第8840号<br>医博第2343号<br>新制||医||764(附属図書館)<br>UT51-2001-F170<br>京都大学大学院医学研究科脳統御医科学系専攻<br>(主査)教授 川口 三郎, 教授 金子 武嗣, 教授 成宮 周<br>学位規則第4条第1項該当

School shootings and emergencies have created the need for educators to be proficient in emergency response procedures; yet they do not always receive the requisite training. The lack of an established delineated training program for New Jersey, kindergarten to Grade 12 institutions has created a situation where educator preparedness varies immensely at schools. Numerous national events of targeted school violence have exemplified the need for quick and proper responses by educators to mitigate the tragic results until first responders arrive. The purpose and goal of this study was to assess educators’ perceptions and to determine the best practices in creating a comprehensive safety and security training program to prepare educators for school crises in order to offer a model for stakeholders to follow or gain ideas to improve their institution’s specific school safety and security emergency plans. Guided by the U.S. Department of Education’s best practices in developing high-quality school safety plans, this study analyzed the perceptions of 60 educators in one New Jersey kindergarten to Grade 12 school on the effectiveness of training. A mixed-methods approach, using a survey questionnaire and interviews, measured changes in the perceptions of these educators after the 15-week program. Data results included a revelation of the implementation of a comprehensive school safety and security plan with related training program resulted in a statistically significant increase in the perceptions of educators’ knowledge and abilities to respond effectively to school targeted violence and emergencies. These findings support the concept that best practices in the field of school safety and security management include appropriate and comprehensive school safety and security plans and training for educators to combat and mitigate school targeted violence and emergent events.

A degradação da qualidade de água no Brasil é um problema sério que afeta grande parte dos rios e lagos. O objetivo desse trabalho é discutir quais mecanismos podem ser usados para a melhora da qualidade das águas tendo em vista a garantia dos seus usos. A legislação brasileira prevê que o sistema de gestão de recursos hídricos deve definir os usos pretendidos para as águas das bacias hidrográficas. Nos casos em que a qualidade das águas precisa ser melhorada para garantir os usos pretendidos, o sistema de gestão de recursos hídricos deve estabelecer etapas progressivas, em que, para cada etapa, são definidas metas de qualidade de água específicas. Utilizando como exemplo o sistema de gestão de qualidade de água de diferentes países, essa dissertação analisa o sistema brasileiro, a definição de metas e a sua relação com o sistema de gestão de recursos hídricos, conforme as definições previstas na Resolução CONAMA 357/05, e identifica os desafios e estratégias para superá-los. Também demonstra que, para que haja mudanças efetivas no cenário de qualidade das águas, será necessário para o país um planejamento estratégico, com prioridades definidas de acordo com as especificidades locais, os investimentos necessários e os aspectos econômicos, enfatizando-se o planejamento e o controle dos serviços de saneamento.<br>Water quality degradation is a serious problem that affects large extensions of rivers and lakes. The purpose of this thesis is to discuss which mechanism can be used to improve water quality in order to guarantee designated uses. The Brazilian water law establishes that the water resource management system must define the designated uses for the watershed. In cases where water quality must be improved to guarantee such uses, the water resource management system establishes a step-by-step system in which, for each step, specific water quality targets are defined. Using as an example the water quality management system of different countries, this thesis analyses the Brazilian system, the target definitions and its relations with the water resource management system, as defined by CONAMA Resolution 357/05, and identifies the challenges and the strategic seams to surpass them. It also demonstrates that, in order to get an effective change in the water quality scenario, it will be necessary for the country to work on strategic planning, with priorities based not only on specific local characteristics, but also on financial needs and economical aspects, with special emphasis on regulation and control of wastewater systems.

Cells invest a lot of energy in order to get their proteins to fold correctly and attain functionality. It is the functional proteome of a cell that defines the ‘life of a cell’. Cells have therefore employed dedicated machinery called chaperones to enable protein folding. One class of these chaperones is heat shock proteins named so because they were initially discovered to be heat inducible and particularly important during heat stress. However the role of heat shock proteins has now been extended from merely being important for stress tolerance. Heat shock proteins are prominently involved in maintaining the correct folding and conformation of proteins and are vital in regulating the stability between protein synthesis and degradation. One of the heat shock proteins, Hsp90, is an evolutionarily conserved molecular chaperone essential in all known eukaryotes examined so far. Unlike other chaperones, Hsp90 is unique in binding to substrate proteins, which are at a late stage of folding, poised for activation by either ligand binding or interaction with other cellular factors. The most common clients of Hsp90 are signaling proteins, the classic example being steroid hormone receptors and signaling kinases. Several other proteins including transcription factors, proteins involved in cell division and development have also been shown to rely on Hsp90 functioning for their maturation. Hsp90 has emerged as an important molecular chaperone due to the large number of proteins that depend on the activity of Hsp90 for their functionality. Hsp90 plays a central role in multiple cellular processes. Since knock-out of hsp90 is lethal to most eukaryotes, inhibitors of Hsp90 have been widely used to study its function. The most widely used inhibitor is geldanamycin (GA). GA binds to the N-terminal/ATP binding site of Hsp90 which results in the degradation of client proteins. Hsp90 clients have been shown to be proteins important for diverse cellular processes such as protein trafficking, signal transduction, cell-cycle, cellular motility and development in eukaryotes. Exploring new Hsp90 clients gives an insight into more pathways that Hsp90 regulates. Intriguingly, many proteins interact with Hsp90 in a context dependent manner, i.e., under certain environmental cue, or in a particular tissue, or only under certain diseased states. It is therefore essential to study Hsp90 functioning and examine Hsp90-client interactions in more than one model organism. Dictyostelium discoideum: a model organism to study the role of Hsp90 in development The eukaryote, Saccharomyces cerevisiae that has been explored extensively for studying the diverse clientele of Hsp90, lacks various signaling pathways important for growth and differentiation as prevalent in higher eukaryotes. It is desirable to develop a model system that would combine the advantages of a lower eukaryote, in terms of its ease of manipulation and retain the complexities of higher eukaryotes. With this motivation, the social slime mold D. discoideum was explored to examine potential roles of cytoplasmic Hsp90 in growth and development. D. discoideum is ideal for studying signaling pathways important for growth and differentiation and to understand how these pathways control cellular responses to external stimuli. Multicellular development in D. discoideum occurs in response to starvation induced stress. As in case of many other protozoans, we conjectured that Hsp90 may participate in regulating developmental transition from unicellular to multicellular stages in Dictyostelium as well. My initial study attempts, to address the role of Hsp90 (HspD), in development of D. discoideum. Towards this two approaches were taken: through genetic interference of HspD, and the other, through its pharmacological inhibition. An antisense HspD plasmid was designed which upon transfection in D. discoideum, showed a very slow growth phenotype, and the cells did not survive beyond few generations. Therefore to further study the functions of HspD, I resorted to pharmacological inhibition by using the specific, well characterized inhibitor, GA. As a first step towards this I examined whether GA was capable of binding to HspD from D. discoideum cell lysate. Towards this, GA was immobilized to NHS-sepharose beads, and bound proteins were examined. Western blot of the bound fraction, using antibody specific to HspD, identified it as a predominant protein being pulled down. This was further confirmed by mass spectrometry. To be able to compare Hsp90 from D. discoideum with Hsp90s from other model organisms, HspD was cloned, purified and biochemically characterized. Comparison of ATPase activities of HspD with Hsp90’s from other systems indicates HspD to possess a relatively low ATPase activity with a Kcat of 1.6 x 10-3 min-1. The dissociation constant of GA for HspD was found to be 0.8 µM, which was in the range similar to Hsp90s from other systems. In addition, we have now obtained structural data on HspD in collaboration with crystallography groups. The N-terminal domain of HspD has been crystallized, both in -free and ligand-bound forms. Crystal structure comparison of HspD with Hsp90 from S. cerevisiae shows overall fold similarity yet some important differences in side chain orientations of specific residues in the ATP binding domain. Interestingly, on treating D. discoideum cells with GA or another Hsp90 N-terminal inhibitor, Radicicol, it was found that, while control cells progressed to develop into fruiting bodies, GA/Radicicol treated cells resulted in delayed development, and were finally arrested at the ‘mound’ stage. This suggested potential involvement of HspD in developmental progression beyond the mound stage. In order to identify the pathways that are probably affected by HspD in D. discoideum development, cells were treated with/without GA and subjected to comparative proteomics using mass spectrometric analysis. Amongst other differences, there was an obvious absence of peptides corresponding to the protein paxillin in GA treated cells. The results were verified by Western blot analysis, using a specific antibody against paxillin, wherein a drastic decrease in paxillin levels were observed in cells treated with GA. Paxillin is a key player in focal adhesion sites that functions as an adaptor protein to recruit diverse cytoskeletal and signaling proteins into a complex, and is essential for cellular proliferation and cell-substrate adhesion. My studies suggest that one of the pathways through which HspD regulates development is through cellular motility as Hsp90 was involved in regulating proteins necessary for motility and cytoskeletal organization at focal adhesion points during development in D. discoideum. Hsp90 as a target for Trypanosoma evansi infections In addition to examining the role of Hsp90 in differentiation in D. discoideum, I have also looked at the potential of Hsp90 under diseased conditions. Towards this, I explored the protozoan parasite, T. evansi, which causes a fatal disease ‘surra’. Surra is a neglected disease that mainly affects domestic and wild animals including equines, camels, cattle and buffaloes. The parasite causes significant economic losses to livestock industry. While this infection is mainly restricted to domestic (camels, equines, cattle, buffaloes, goats, sheep, pigs, dogs etc.) and wild animals, recent reports indicate their ability to infect humans. There are no reliable sensitive and specific diagnostic tests or vaccines available against this disease and the available drugs show significant toxicity. There is an urgent need to develop improved methods of diagnosis and control measures for this disease. Unlike its related human parasites T. brucei and T. cruzi whose genomes have been fully sequenced T. evansi genome sequence remains unavailable. With a view to identifying potential diagnostic markers and drug targets I have studied the clinical proteome of T. evansi infection using mass spectrometry. I have been able to identify almost 166 proteins of T. evansi, which also included potential drug and vaccine targets. Due to absence of any genome sequence information from T. evansi, most of the peptides obtained matched to its related species, T. brucei, T. cruzi and also few from Leishmania major. Importantly, I was also able to identify peptides from Hsp90. Hsp90 from T. evansi was cloned and its sequence was also obtained. To investigate the possibility of exploring Hsp90 as a target against Surra infections, TeHsp90 protein was purified by expressing it in bacterial cells, and its drug (GA) binding ability was examined in-vitro. The dissociation constant of GA for HspD was found to be 1.4 µM, which was in the range similar to Hsp90s from other systems. The ability of 17AAG (a derivative of GA) was examined in inhibiting T. evansi infection at pre-clinical level. Towards this, swiss female mice were infected with purified parasites and then the drug was injected either immediately, in one group of mice, and in another group of mice the parasites were challenged with the drug only after the onset of infection. Interestingly, both groups of mice were found to get cured using Hsp90 inhibitor. The pre-clinical results suggested that Hsp90 was an interesting drug target and its inhibitor could indeed be used against ‘surra’ infections. Hsp90 from Giardia lamblia: An unusual case Hsp90 was also examined from another pathogenic protozoan, Giardia lamblia, one of the leading causes of diarrhea in the world. Previous studies from our lab have shown Gardial Hsp90 to be coded by two different ORFs, spliced together in trans. This is indeed the only example of trans-splicing in Hsp90 known so far. My study further characterizes this finding through analysis of transcription levels of the individual ORFs, using Northern blot analysis. Importantly, I was able to detect transcripts of all three forms of Hsp90; full-length, N terminus as well as C terminus, suggesting that these are expressed and may have biological significance. To understand the significance of these independent transcripts, I have examined relative levels of expression of all three forms by Real-time PCR analysis wherein there was almost 90 fold and 5 fold lesser transcript level of N terminus and C terminus Hsp90 observed, respectively as compared to the full-length GlHsp90 expression. Previous reports have shown Hsp90 from all known organisms, to get up regulated during heat shock. Thus it was important to examine the effect of heat stress on the expression of these independent transcripts. Interestingly, different domains were found to get independently induced during heat stress. The transcript level of HspC was seen to be almost similar to that of full-length upon heat shock. There was also a significant up regulation observed in HspN transcript upon heat shock. Taking together all these observations, these results suggest a possible role for the independent domains, HspN and HspC during heat stress in G. lamblia. Furthermore, I have cloned and purified one of the individually expressed domains, HspN and characterized it biochemically. HspN was found to be able to bind to ATP, however lacked ATPase activity. Taking together all these observations, it suggests a possible role for the independent domains, HspN and HspC which needs to be investigated further. Summary Altogether, my studies establish the importance of alternate model systems in understanding the biology of Hsp90. The importance of Hsp90 was first established in growth and development of a nonpathogenic protozoan D. discoideum. My results provide significant insights into the additional pathways that Hsp90 regulates during D. discoideum development. One such important pathway was delineated to be cellular locomotion and motility. Further, I have also studied the importance of Hsp90 in neglected infectious diseases. In addition to providing a glimpse into the pathways operational during disease manifestation in T. evansi, we have shown Hsp90 to be effective in pre-clinical trials against T. evansi infections. Hsp90 from another pathogenic protozoan, G. lamblia, has also been studied. This is by far the only organism, in which there is an independent expression of the N-and C-terminal domain of Hsp90. The rare gene organization, coupled with independent expression of domains of Hsp90, makes this organism important to examine novel functions of this chaperone.

Cells invest a lot of energy in order to get their proteins to fold correctly and attain functionality. It is the functional proteome of a cell that defines the ‘life of a cell’. Cells have therefore employed dedicated machinery called chaperones to enable protein folding. One class of these chaperones is heat shock proteins named so because they were initially discovered to be heat inducible and particularly important during heat stress. However the role of heat shock proteins has now been extended from merely being important for stress tolerance. Heat shock proteins are prominently involved in maintaining the correct folding and conformation of proteins and are vital in regulating the stability between protein synthesis and degradation. One of the heat shock proteins, Hsp90, is an evolutionarily conserved molecular chaperone essential in all known eukaryotes examined so far. Unlike other chaperones, Hsp90 is unique in binding to substrate proteins, which are at a late stage of folding, poised for activation by either ligand binding or interaction with other cellular factors. The most common clients of Hsp90 are signaling proteins, the classic example being steroid hormone receptors and signaling kinases. Several other proteins including transcription factors, proteins involved in cell division and development have also been shown to rely on Hsp90 functioning for their maturation. Hsp90 has emerged as an important molecular chaperone due to the large number of proteins that depend on the activity of Hsp90 for their functionality. Hsp90 plays a central role in multiple cellular processes. Since knock-out of hsp90 is lethal to most eukaryotes, inhibitors of Hsp90 have been widely used to study its function. The most widely used inhibitor is geldanamycin (GA). GA binds to the N-terminal/ATP binding site of Hsp90 which results in the degradation of client proteins. Hsp90 clients have been shown to be proteins important for diverse cellular processes such as protein trafficking, signal transduction, cell-cycle, cellular motility and development in eukaryotes. Exploring new Hsp90 clients gives an insight into more pathways that Hsp90 regulates. Intriguingly, many proteins interact with Hsp90 in a context dependent manner, i.e., under certain environmental cue, or in a particular tissue, or only under certain diseased states. It is therefore essential to study Hsp90 functioning and examine Hsp90-client interactions in more than one model organism. Dictyostelium discoideum: a model organism to study the role of Hsp90 in development The eukaryote, Saccharomyces cerevisiae that has been explored extensively for studying the diverse clientele of Hsp90, lacks various signaling pathways important for growth and differentiation as prevalent in higher eukaryotes. It is desirable to develop a model system that would combine the advantages of a lower eukaryote, in terms of its ease of manipulation and retain the complexities of higher eukaryotes. With this motivation, the social slime mold D. discoideum was explored to examine potential roles of cytoplasmic Hsp90 in growth and development. D. discoideum is ideal for studying signaling pathways important for growth and differentiation and to understand how these pathways control cellular responses to external stimuli. Multicellular development in D. discoideum occurs in response to starvation induced stress. As in case of many other protozoans, we conjectured that Hsp90 may participate in regulating developmental transition from unicellular to multicellular stages in Dictyostelium as well. My initial study attempts, to address the role of Hsp90 (HspD), in development of D. discoideum. Towards this two approaches were taken: through genetic interference of HspD, and the other, through its pharmacological inhibition. An antisense HspD plasmid was designed which upon transfection in D. discoideum, showed a very slow growth phenotype, and the cells did not survive beyond few generations. Therefore to further study the functions of HspD, I resorted to pharmacological inhibition by using the specific, well characterized inhibitor, GA. As a first step towards this I examined whether GA was capable of binding to HspD from D. discoideum cell lysate. Towards this, GA was immobilized to NHS-sepharose beads, and bound proteins were examined. Western blot of the bound fraction, using antibody specific to HspD, identified it as a predominant protein being pulled down. This was further confirmed by mass spectrometry. To be able to compare Hsp90 from D. discoideum with Hsp90s from other model organisms, HspD was cloned, purified and biochemically characterized. Comparison of ATPase activities of HspD with Hsp90’s from other systems indicates HspD to possess a relatively low ATPase activity with a Kcat of 1.6 x 10-3 min-1. The dissociation constant of GA for HspD was found to be 0.8 µM, which was in the range similar to Hsp90s from other systems. In addition, we have now obtained structural data on HspD in collaboration with crystallography groups. The N-terminal domain of HspD has been crystallized, both in -free and ligand-bound forms. Crystal structure comparison of HspD with Hsp90 from S. cerevisiae shows overall fold similarity yet some important differences in side chain orientations of specific residues in the ATP binding domain. Interestingly, on treating D. discoideum cells with GA or another Hsp90 N-terminal inhibitor, Radicicol, it was found that, while control cells progressed to develop into fruiting bodies, GA/Radicicol treated cells resulted in delayed development, and were finally arrested at the ‘mound’ stage. This suggested potential involvement of HspD in developmental progression beyond the mound stage. In order to identify the pathways that are probably affected by HspD in D. discoideum development, cells were treated with/without GA and subjected to comparative proteomics using mass spectrometric analysis. Amongst other differences, there was an obvious absence of peptides corresponding to the protein paxillin in GA treated cells. The results were verified by Western blot analysis, using a specific antibody against paxillin, wherein a drastic decrease in paxillin levels were observed in cells treated with GA. Paxillin is a key player in focal adhesion sites that functions as an adaptor protein to recruit diverse cytoskeletal and signaling proteins into a complex, and is essential for cellular proliferation and cell-substrate adhesion. My studies suggest that one of the pathways through which HspD regulates development is through cellular motility as Hsp90 was involved in regulating proteins necessary for motility and cytoskeletal organization at focal adhesion points during development in D. discoideum. Hsp90 as a target for Trypanosoma evansi infections In addition to examining the role of Hsp90 in differentiation in D. discoideum, I have also looked at the potential of Hsp90 under diseased conditions. Towards this, I explored the protozoan parasite, T. evansi, which causes a fatal disease ‘surra’. Surra is a neglected disease that mainly affects domestic and wild animals including equines, camels, cattle and buffaloes. The parasite causes significant economic losses to livestock industry. While this infection is mainly restricted to domestic (camels, equines, cattle, buffaloes, goats, sheep, pigs, dogs etc.) and wild animals, recent reports indicate their ability to infect humans. There are no reliable sensitive and specific diagnostic tests or vaccines available against this disease and the available drugs show significant toxicity. There is an urgent need to develop improved methods of diagnosis and control measures for this disease. Unlike its related human parasites T. brucei and T. cruzi whose genomes have been fully sequenced T. evansi genome sequence remains unavailable. With a view to identifying potential diagnostic markers and drug targets I have studied the clinical proteome of T. evansi infection using mass spectrometry. I have been able to identify almost 166 proteins of T. evansi, which also included potential drug and vaccine targets. Due to absence of any genome sequence information from T. evansi, most of the peptides obtained matched to its related species, T. brucei, T. cruzi and also few from Leishmania major. Importantly, I was also able to identify peptides from Hsp90. Hsp90 from T. evansi was cloned and its sequence was also obtained. To investigate the possibility of exploring Hsp90 as a target against Surra infections, TeHsp90 protein was purified by expressing it in bacterial cells, and its drug (GA) binding ability was examined in-vitro. The dissociation constant of GA for HspD was found to be 1.4 µM, which was in the range similar to Hsp90s from other systems. The ability of 17AAG (a derivative of GA) was examined in inhibiting T. evansi infection at pre-clinical level. Towards this, swiss female mice were infected with purified parasites and then the drug was injected either immediately, in one group of mice, and in another group of mice the parasites were challenged with the drug only after the onset of infection. Interestingly, both groups of mice were found to get cured using Hsp90 inhibitor. The pre-clinical results suggested that Hsp90 was an interesting drug target and its inhibitor could indeed be used against ‘surra’ infections. Hsp90 from Giardia lamblia: An unusual case Hsp90 was also examined from another pathogenic protozoan, Giardia lamblia, one of the leading causes of diarrhea in the world. Previous studies from our lab have shown Gardial Hsp90 to be coded by two different ORFs, spliced together in trans. This is indeed the only example of trans-splicing in Hsp90 known so far. My study further characterizes this finding through analysis of transcription levels of the individual ORFs, using Northern blot analysis. Importantly, I was able to detect transcripts of all three forms of Hsp90; full-length, N terminus as well as C terminus, suggesting that these are expressed and may have biological significance. To understand the significance of these independent transcripts, I have examined relative levels of expression of all three forms by Real-time PCR analysis wherein there was almost 90 fold and 5 fold lesser transcript level of N terminus and C terminus Hsp90 observed, respectively as compared to the full-length GlHsp90 expression. Previous reports have shown Hsp90 from all known organisms, to get up regulated during heat shock. Thus it was important to examine the effect of heat stress on the expression of these independent transcripts. Interestingly, different domains were found to get independently induced during heat stress. The transcript level of HspC was seen to be almost similar to that of full-length upon heat shock. There was also a significant up regulation observed in HspN transcript upon heat shock. Taking together all these observations, these results suggest a possible role for the independent domains, HspN and HspC during heat stress in G. lamblia. Furthermore, I have cloned and purified one of the individually expressed domains, HspN and characterized it biochemically. HspN was found to be able to bind to ATP, however lacked ATPase activity. Taking together all these observations, it suggests a possible role for the independent domains, HspN and HspC which needs to be investigated further. Summary Altogether, my studies establish the importance of alternate model systems in understanding the biology of Hsp90. The importance of Hsp90 was first established in growth and development of a nonpathogenic protozoan D. discoideum. My results provide significant insights into the additional pathways that Hsp90 regulates during D. discoideum development. One such important pathway was delineated to be cellular locomotion and motility. Further, I have also studied the importance of Hsp90 in neglected infectious diseases. In addition to providing a glimpse into the pathways operational during disease manifestation in T. evansi, we have shown Hsp90 to be effective in pre-clinical trials against T. evansi infections. Hsp90 from another pathogenic protozoan, G. lamblia, has also been studied. This is by far the only organism, in which there is an independent expression of the N-and C-terminal domain of Hsp90. The rare gene organization, coupled with independent expression of domains of Hsp90, makes this organism important to examine novel functions of this chaperone.

碩士<br>長庚大學<br>醫學生物技術暨檢驗學系<br>101<br>Human enterovirus 71 (EV71) genome replication is catalyzed by viral 3D RNA dependent RNA polymerase (RdRP). We identified that 3D polymerase associated with nuclear factor of activated T cell,90-KD (NF90) in EV71 infected cell by matrix-associated laser desorption/ ionization-time of flight (MALDI-TOF) analysis. RNA-binding protein NF90 is an indispensable factor for post-transcriptional regulation and has been implicated in the translational control of target mRNAs which bearing AU-rich signature motif in the 3’UTR. The aim of this study is to elucidate viral 3D polymerase-NF90 interaction that interfere host mRNAs expression and EV71 viral replication. First, we performed confocal microscopy and co-immunoprecipitation to prove that NF90 colocalized with 3D polymerase in cytosol upon EV71 infection and the association between NF90 and 3D polymerase required RNA as a intermediate. Furthermore, We utilized biotinylated RNA-protein pull-down assay to discover that 3D polymerase recombinant protein disrupts the association of NF90 and target mRNAs. On the other hand, we observed NF90 suppress viral replication at early time post infection both in EV71-FLuc replicon transfected and virus infected cells. Taken together, the study demonstrates that EV71 3D polymerase impairs NF90-target mRNAs interaction. Moreover, we also obtained evidence that NF90 suppresses EV71 IRES-driven translation at early time post infection.

The ferroelectric domains of thin-film lithium niobate are structured for multi-wavelength nonlinear optical frequency conversion. We target frequency-multiplexed photon-pair generation, in which we show conditions for five pair sources (purity &gt; 90%) in a single waveguide.

Bangladesh has made remarkable progress in achieving over 90% primary enrollment with significantly improved gender parity. Its focus has now shifted from universal primary education towards the quality of education and improvements in teaching outcomes. In order to meet this goal, the government has turned to the merits that EdTech can offer and is implementing a blended learning approach that integrates both digital and physical elements, and balances creative and logical aspects to strengthen its progress and address Bangladesh’s current educational challenges. In partnership with EdTech Hub, this Spotlight contributes to identifying EdTech innovations that have a great potential to be scaled and are being impactfully implemented in the context of Bangladesh and beyond, to directly address and support the learning needs of children, as well as supporting education stakeholders such as educators, parents, and communities coming from varying social and economic backgrounds. We sought a range of hi- to low-tech solutions, encompassing everything from SMS-based technologies and game-based learning to school management systems. These innovations target various educational stakeholders, including teachers, students, and administrators. This Spotlight also demonstrates how private and public sector players fit in the EdTech innovation ecosystem.

The aviation sector has a significant role to play in the reduction of carbon emissions, particularly in light of EU emissions targets. Within the EU, aviation emissions constitute 13.9 per cent of total transport emissions, where under the Green Deal a target of 90 per cent reduction in transport emissions by 2050 has been set. Several tax exemptions apply to the aviation sector due to its international nature, which have contributed to an increasing trend in emissions. Recent movements in EU policy under the ‘Fit for 55’ package, as well as public and political interest, have focused attention on changing the taxation struc-ture of the aviation industry in order to reduce CO2 emissions. This report focuses on several different potential policies and taxation structures which would attempt to decrease the emissions of the aviation sector: the removal of kerosene taxation exemption (in line with the recent EU proposal), removal of VAT exemption, introduction of a passenger tax, and abolition of free EU ETS allowances by 2026 (in line with the recent EU proposal). Each of these policies, as well a combination scenario, is assessed with respect to the impacts on the aviation industry, spillover effects, macro-economic and government revenue effects, and household effects, as well as the emissions impacts. This analysis applies the Ireland, Environment, Energy and Economy (I3E) model, which allows for the examination of spillover effects of the aviation industry to government, households and other industries. This methodology does not, however, allow for a detailed analysis of different passenger or airline types. In what follows, results are shown as cumulative percentage changes compared to no aviation taxation. Hence a decrease does not present an annual decrease but the build up of a cumulative decrease from 2022 to the given year.

The Country Diagnostic Study (CDS) for Senegal uses the Hausmann-Rodrik-Velasco growth diagnostics model to identify the binding constraints being faced in its quest for higher and more sustained economic growth and make recommendations to relax these constraints. Hence, the findings of the CDS can help the Islamic Development Bank in identifying areas where it can have a greater impact and provide an evidence-basis to support the development of the Member Country Partnership Strategy. After decades of subdued and highly volatile economic growth due to heavy dependence on primary commodities and low productivity, Senegal experienced an unprecedented growth acceleration from 2014 to 2019. However, there appeared to be a weak correlation between economic growth and jobs creation. In addition, about 90 percent of non-agricultural employment is estimated to be informal. The national poverty rate decreased by 5 percentage points between 2011 and 2018. Nonetheless, the absolute number of poor people has increased. Furthermore, regional disparities are persistent. Despite the country’s solid performance in the field of governance, further simplification and transparency of business procedures and regulations will be critical in addressing the challenge of informality. Efforts to address informality in the economy should also target the issue of access to finance through the design of financing mechanisms based on specific needs assessment and risk management tools. Senegal will also need to create the conditions for higher competitiveness and follow upgrading trajectories in global and regional value chains. In this respect, both physical and digital connectivity will be essential.

The proposed project aims to enhance tree fruit identification and targeting for robotic harvesting through the selection of appropriate sensor technology, sensor fusion, and visual servo-control approaches. These technologies will be applicable for apple, orange and grapefruit harvest, although specific sensor wavelengths may vary. The primary challenges are fruit occlusion, light variability, peel color variation with maturity, range to target, and computational requirements of image processing algorithms. There are four major development tasks in original three-year proposed study. First, spectral characteristics in the VIS/NIR (0.4-1.0 micron) will be used in conjunction with thermal data to provide accurate and robust detection of fruit in the tree canopy. Hyper-spectral image pairs will be combined to provide automatic stereo matching for accurate 3D position. Secondly, VIS/NIR/FIR (0.4-15.0 micron) spectral sensor technology will be evaluated for potential in-field on-the-tree grading of surface defect, maturity and size for selective fruit harvest. Thirdly, new adaptive Lyapunov-basedHBVS (homography-based visual servo) methods to compensate for camera uncertainty, distortion effects, and provide range to target from a single camera will be developed, simulated, and implemented on a camera testbed to prove concept. HBVS methods coupled with imagespace navigation will be implemented to provide robust target tracking. And finally, harvesting test will be conducted on the developed technologies using the University of Florida harvesting manipulator test bed. During the course of the project it was determined that the second objective was overly ambitious for the project period and effort was directed toward the other objectives. The results reflect the synergistic efforts of the three principals. The USA team has focused on citrus based approaches while the Israeli counterpart has focused on apples. The USA team has improved visual servo control through the use of a statistical-based range estimate and homography. The results have been promising as long as the target is visible. In addition, the USA team has developed improved fruit detection algorithms that are robust under light variation and can localize fruit centers for partially occluded fruit. Additionally, algorithms have been developed to fuse thermal and visible spectrum image prior to segmentation in order to evaluate the potential improvements in fruit detection. Lastly, the USA team has developed a multispectral detection approach which demonstrated fruit detection levels above 90% of non-occluded fruit. The Israel team has focused on image registration and statistical based fruit detection with post-segmentation fusion. The results of all programs have shown significant progress with increased levels of fruit detection over prior art.

2016 will surely be the hottest year since records began in the 19th century. The increase will be very close to the target set in the Paris Agreement to avoid an increase in global temperature by 1.5 °C. Average temperaturesin 2016 have risen to 1.2 °C above what they were before the industrial revolution. The dilemma facing the world today, in view of these data, becomes even more urgent: How to reduce greenhouse gas emissions from fossil fuels, accepting that their demand will continue to exist in the coming decades? In the energy sector, many solutions have been proposed to completely replace fossil fuels for electricity generation, such as massive deployment of renewable energy generation and increased energy efficiency. There are many restrictions, however, to achieve this result in the medium term, ranging from technological limitations in the massive deployment of energy efficiency and renewable energies, to the political economy of countries that are unlikely to reduce their oil production And carboniferous as long as demand exists. Carbon capture and storage (CCS) offers an alternative to mitigate CO2 emissions from fossil fuel power plants, considering that, given current and future energy needs, the operation of these plants will continue in the coming years. CAC could mitigate up to 90% of the carbon dioxide emitted by the use of fossil fuels in electricity generation and industrial processes. Additionally, the use of CAC with renewable biomass is one of the few carbon reduction technologies that can be used in a "carbon-negative" mode. If biomass from fuelwood crops were used, carbon could be absorbed and simultaneously generate electricity. CCS, therefore, is a viable alternative to solve the dilemma of reducing emissions while satisfying the growing energy needs of the world.

Expansion of piped water and sewage collection is a critical issue in developing countries, where public investment is limited. In Brazil, the New Sanitation Regulatory Framework from 2020 intends to stimulate the entry of private providers into the market while setting connection targets: 99% of households with piped water and 90%with piped sewer. We assess the viability of these connection goals under the current regulated prices and compare the distributional impacts of supplemental policies. We use billing data from a major private provider and a structural model encompassing the expansion decisions of the service provider and the connection and consumption choices of consumers. We find that even if the firm expanded water and sewer everywhere, the targets would not be met, with only 43% connecting to piped sewage. Moreover, the expansion is not profitable for the firm. We then simulate charging consumers when piped sewage is available, regardless of connection, and price discounts on the sewer bills. Combinations of these two policies stimulate the firm to expand and consumers to connect, achieving 76% connected to sewer but reducing consumer surplus.

In spite of the fact that they make up more than 90% of firms and account for a substantial portion of employment in these countries, rather little is known about the role of small- and mid-size enterprises (SMEs) in Latin American and the Caribbean (LAC) trade. In particular, there are a series of pending yet pivotal questions, such as: To what extent and how do SMEs contribute to their countries exports? How internationalized are LAC SMEs compared to SMEs in other world regions? How diversified are LAC SMEs exports across products and destinations? What are the key bottlenecks to LAC SME internationalization? This issue of the Journal intends to drive a holistic, thoughtful and targeted discussion of patterns of SME internationalization and its major bottlenecks and how they can be (or not) mitigated most effectively.

In 1992 a working group of the UK Advisory Committee on the Microbiological Safety of Food presented a report on Vacuum Packaging and Associated Processes regarding the microbiological safety of chilled foods. The report supported subsequent guidance provided by the UK Food Standards Agency for the safe manufacture of vacuum packed and modified atmosphere packed chilled foods. In 2021 the ACMSF requested that a new subgroup should update and build on the 1992 report as well as considering, in addition to chilled foods, some foods that are intended to be stored at ambient temperatures. The new subgroup agreed a scope that includes the conditions that support growth and/or neurotoxin formation by C. botulinum, and other clostridia, as well as identification of limiting conditions that provide control. Other foodborne pathogens that need to be considered separately and some foods including raw beef, pork and lamb were explicitly excluded. The subgroup considered the taxonomy, detection, epidemiology, occurrence, growth, survival and risks associated with C. botulinum and other neurotoxin-forming clostridia. There has been no significant change in the nature of foodborne botulism in recent decades except for the identification of rare cases caused by neurotoxigenic C. butyricum, C. baratii and C. sporogenes. Currently evidence indicates that non-clostridia do not pose a risk in relation to foodborne botulism. The subgroup has compiled lists of incidents and outbreaks of botulism, reported in the UK and worldwide, and have reviewed published information concerning growth parameters and control factors in relation to proteolytic C. botulinum, non-proteolytic C. botulinum and the other neurotoxigenic clostridia. The subgroup concluded that the frequency of occurrence of foodborne botulism is very low (very rare but cannot be excluded) with high severity (severe illness: causing life threatening or substantial sequelae or long-term illness). Uncertainty associated with the assessment of the frequency of occurrence, and with the assessment of severity, of foodborne botulism is low (solid and complete data; strong evidence in multiple sources). The vast majority of reported botulism outbreaks, for chilled or ambient stored foods, are identified with proteolytic C. botulinum and temperature abuse is the single most common cause. In the last 30 years, in the UK and worldwide where a cause can be identified, there is evidence that known controls, combined with the correct storage, would have prevented the reported incidents of foodborne botulism. The subgroup recommends that foods should continue to be formulated to control C. botulinum, and other botulinum neurotoxin-producing clostridia, in accordance with the known factors. With regard to these controls, the subgroup recommends some changes to the FSA guidelines that reflect improved information about using combinations of controls, the z-value used to establish equivalent thermal processes and the variable efficacy associated with some controls such as herbs and spices. Current information does not facilitate revision of the current reference process, heating at 90°C for 10 minutes, but there is strong evidence that this provides a lethality that exceeds the target 6 order of magnitude reduction in population size that is widely attributed to the process and the subgroup includes a recommendation that the FSA considers this issue. Early detection and connection of cases and rapid, effective coordinated responses to very rare incidents are identified as crucial elements for reducing risks from foodborne botulism. The subgroup recommends that the FSA works closely with other agencies to establish clear and validated preparedness in relation to potential major incidents of foodborne botulism in the UK.

Freshwater habitats, such as rivers and wetlands, provide major benefits to society through services such as flood risk reduction and drinking water. However, many of these habitats are in a poor condition. They have been damaged by a range of human activities including physical modification and pollution. For example, an estimated 90% of wetlands have been lost over the last century. Restoration of freshwater habitats can be achieved by addressing the causes of degradation and by enhancing or extending habitats. There is good evidence on the benefits of some forms of restoration. For example, restoration can deliver effective flood defence and restore fish populations. However, certain restoration measures are less well-studied. There are numerous national and international targets for the restoration of freshwater habitats. These include a commitment to improving at least 75% of waters to close to their natural state as soon as practicable by 2042. However, some stakeholders believe that these commitments will not be met under current plans in England. There have been calls for various actions including: increased and long-term funding to deliver larger-scale projects changes to policy and legislation to deliver more joined-up decision making and to include small water bodies and headwaters further prevention of degradation, including better enforcement of existing legislation the removal of barriers to nature-based solutions Policy in this area is devolved. For England, the UK Government published a Plan for Water in 2023 that contained new actions to help improve the condition of freshwater habitats. The Office for Environmental Protection will publish an assessment of the Government’s approach in 2024.