Novel Flavin Chemistry Involved in the Biosynthesis of the Lower Ligand of Vitamin B12

Vitamin B₁₂ or Cobalamin is structurally the most complex of all the vitamins. The entire biosynthetic pathway of cobalamin is well studied except the formation of the lower ligand known as dimethylbenzimidazole (DMB). In aerobic pathway, BluB catalyzes the formation of DMB using reduced flavin mononucleotide (FMNH₂) as its substrate leading to the formation of DMB and erythrose4-phosphate as the end products. It is a remarkable transformation, wherein, C2 of DMB is derived from C1' of the ribose sugar chain of FMN. However, the fate of the ring C of FMN moiety as well as the overall mechanism of this unique reaction is still unknown. Thus, identification of the final unknown product and a detailed mechanistic study of dimethylbenzimidazole formation is the main focus of this work. In this dissertation, we have successfully identified alloxan as the end product derived from the third ring of FMN. Water and molecular oxygen have been shown to be the two sources of oxygen atom incorporation in alloxan based on O-18 labeling studies. A key intermediate in our mechanistic proposal has been successfully trapped in the form of six different shunt products using water, bisulfite and hydride as nucleophiles. Trapping of this intermediate helps us establish that the C-C bond cleavage occurs first forming erythrose-4-phosphate followed by the release of alloxan. Asp-32 has been shown to play an important role in stabilizing this intermediate. Based on stereochemical studies, we have shown that the pro-R hydrogen is selectively abstracted from the C1' of the ribityl chain of the substrate. This result helps us establish that the final oxidation step involved in DMB formation is indeed catalyzed by the enzyme. Formation of a new shunt product was observed on using 8-substituted flavins as substrate analogs for the BluB catalyzed reaction. Characterization of this shunt product provides evidence for the initial fragmentation of the peroxyflavin intermediate involved in the DMB biosynthesis. All these above observations are consistent with our current mechanistic proposal for the DMB formation. Thus, we have finally unraveled the long unsolved mystery in the Vitamin B₁₂ biosynthesis

Novel Flavin Chemistry Involved in the Biosynthesis of the Lower Ligand of Vitamin B12

Vitamin B₁₂ or Cobalamin is structurally the most complex of all the vitamins. The entire biosynthetic pathway of cobalamin is well studied except the formation of the lower ligand known as dimethylbenzimidazole (DMB). In aerobic pathway, BluB catalyzes the formation of DMB using reduced flavin mononucleotide (FMNH₂) as its substrate leading to the formation of DMB and erythrose4-phosphate as the end products. It is a remarkable transformation, wherein, C2 of DMB is derived from C1' of the ribose sugar chain of FMN. However, the fate of the ring C of FMN moiety as well as the overall mechanism of this unique reaction is still unknown. Thus, identification of the final unknown product and a detailed mechanistic study of dimethylbenzimidazole formation is the main focus of this work. In this dissertation, we have successfully identified alloxan as the end product derived from the third ring of FMN. Water and molecular oxygen have been shown to be the two sources of oxygen atom incorporation in alloxan based on O-18 labeling studies. A key intermediate in our mechanistic proposal has been successfully trapped in the form of six different shunt products using water, bisulfite and hydride as nucleophiles. Trapping of this intermediate helps us establish that the C-C bond cleavage occurs first forming erythrose-4-phosphate followed by the release of alloxan. Asp-32 has been shown to play an important role in stabilizing this intermediate. Based on stereochemical studies, we have shown that the pro-R hydrogen is selectively abstracted from the C1' of the ribityl chain of the substrate. This result helps us establish that the final oxidation step involved in DMB formation is indeed catalyzed by the enzyme. Formation of a new shunt product was observed on using 8-substituted flavins as substrate analogs for the BluB catalyzed reaction. Characterization of this shunt product provides evidence for the initial fragmentation of the peroxyflavin intermediate involved in the DMB biosynthesis. All these above observations are consistent with our current mechanistic proposal for the DMB formation. Thus, we have finally unraveled the long unsolved mystery in the Vitamin B₁₂ biosynthesis

LAPORAN INDIVIDU PRAKTIK PENGALAMAN LAPANGAN (PPL) SMA NEGERI 10 YOGYAKARTA Periode 1 Juli – 17 September

Praktek Pengalaman Lapangan (PPL) 2014 dilaksanakan di SMA Negeri 10 Yogyakarta, bertujuan untuk memberikan pengalaman pada mahasiswa dalam bidang pembelajaran dalam rangka melatih dan mengembangkan potensi keguruan atau kependidikan. Praktek Pengalaman Lapangan (PPL) memberikan kesempatan kepada mahasiswa untuk mempelajari, mengenal, dan melaksanakan praktik pembelajaran kelas, serta dapat digunakan oleh mahasiswa untuk meningkatkan kemampuan untuk menerapkan ilmu pengetahuan dan keterampilan yang telah dikuasai secara interdisipliner ke dalam kehidupan nyata di sekolah yaitu dengan cara menumbuhkan motivasi atas dasar kekuatan sendiri. Berdasarkan hasil observasi dan wawancara dengan berbagai pihak, ditemukan beberapa persoalan atau permasalahan yang ada di SMA Negeri 10 Yogyakarta. Melihat situasi dan kondisi tersebut, maka dibentuk suatu rancangan program kerja PPL sebagai upaya menangani permasalahan yang ada. Program Kerja Praktik Pengalaman Lapangan (PPL) yang dirancang meliputi: Rencana Pelaksanaan Pembelajaran (RPP), pembuatan media pembelajaran, pembuatan soal ulangan harian. Secara keseluruhan program kerja PPL dapat terlaksana dengan baik dan lancar walaupun dalam proses pembelajaran nyata, banyak yang menggunakan modifikasi alat. Hasil dari program kerja meliputi: terbuatnya Rencana Pelaksanaan Pembelajaran (RPP), soal ulangan harian, pembuatan daftar nilai siswa, dan media pembelajaran

A Learning Culture and Continuous Learning for a Learning Organization

This paper is aimed at studying a learning culture and continuous learning and finding out the benefits and the ways to develop continuous learning in an organization. The definition of learning culture and continuous learning is inferred from some experts’ opinions such as Van Breda-Verduijn & Heijboer, Cerne Matej, Blackwood, Schein, Jain & Martindale, etc. An organizational learning culture is inferred as a set of norms and values about the functioning of an organization that encourages individuals or the organization to carry out continuous learning. A continuous learning culture is important to implement in an organization since it has many benefits for a learning organization. An organizational learning culture can be developed if there are supports in forms of motivation, learning opportunities, and facilities to its members.   Keywords: Learning Culture, Continuous Learning, Learning Organizatio

Hubungan Estimasi Kadar Digoksin Dalam Darah Dengan Kejadian Toksisitas Digoksin pada Pasien Rawat Inap Bangsal Penyakit Dalam RSUD Prof. Dr. Margono Soekarjo Purwokerto

Latar Belakang: Digoksin merupakan obat dengan indeks terapi sempit untuk penyakit kardiovaskuler sehingga kenaikan atau penurunan sedikit kadar obat dalam darah dapat membuat kadar berada di luar rentang terapi. Oleh sebab itu, diperlukan pemantauan kadar digoksin dalam darah untuk mengurangi resiko toksisitas digoksin, salah satunya menggunakan metode farmakokinetika. Penelitian ini bertujuan untuk mengetahui persentase pasien yang memiliki estimasi kadar digoksin >2 ng/ml, mengetahui tanda – tanda toksisitas yang muncul dan hubungan estimasi kadar digoksin dengan tanda toksisitas yang muncul pada pasien rawat inap bangsal penyakit dalam RSUD Prof. dr. Margono Soekarjo Purwokerto. Metode: Penelitian bersifat prospektif selama 2 bulan melalui pencatatan rekam medis dan wawancara secara langsung kepada pasien. Sampel dipilih dengan cara total sampling sesuai kriteria inklusi dan eksklusi. Data diolah dengan analisis univariat sedangkan untuk hubungan estimasi kadar dengan tanda toksisitas yang muncul dihubungakan menggunakan uji koefisien kontingensi. Hasil: Jumlah pasien dengan estimasi kadar digoksin 2 ng/ml sebanyak 13 pasien (38,2%). Tanda toksisitas muncul pada 6 pasien (17,6%) dimana 3 pasien (8,8%) berada pada rentang ≤2 ng/ml dan 3 pasien (8,8%) berada pada rentang >2 ng/ml. Tanda toksisitas yang paling banyak muncul yaitu perubahan EKG terkait aritmia. Estimasi kadar tunak digoksin dalam darah >2 ng/ml tidak berhubungan langsung secara statistik (p value 0,513) dengan munculnya tanda – tanda toksisitas digoksin. Kesimpulan: Kadar digoksin dalam darah harus dipantau lebih akurat lagi menggunakan sampling darah secara langsung

ANALISIS STRUKTURAL-SEMIOTIK ROMAN LA VIE EST BRÈVE ET LE DÉSIR SANS FIN KARYA PATRICK LAPEYRE

L’objectif de cette recherche sont (1) de décrire les éléments intrinsèques du roman sous forme l’intrigue, les personnages, l’espace, et le thème dans le roman La Vie est Brève et le Désir sans Fin de Patrick Lapeyre, (2) de décrire la relation entre ces éléments intrinsèques, (3) de décrire l’analyse sémiotique du roman sous forme la relation entre le représentamen et le fond, la relation entre le signe et le référent, et la relation entre le signe et l’interprétant dans le roman La Vie est Brève et le Désir sans Fin de Patrick Lapeyre. Le sujet de cette recherche est le roman La Vie est Brève et le Désir sans Fin de Patrick Lapeyre publie en 2010. L’objet de cette recherche est (1) les éléments intrinsèques qui existent dans ce roman sous forme l’intrigue, le personnage, l’espace et le thème, (2) les liens entre ces éléments intrinsèques, (3) la forme de la relation entre le représentamen et le fond, la relation entre le signe et le référent, et la relation entre le signe et l’interprétant dans le roman. La méthode utilisée est la méthode descriptives-qualitative avec la technique d’analyse du contenu. La validité des données est déterminée par la validité de la sémantique. La fiabilité des données utilisées est la fiabilité intra-rater et soutenu par un jugement d'expert. Le résultat de cette recherche montre que (1) le roman La Vie est Brève et le Désir sans Fin de Patrick Lapeyre a une intrigue progressive avec cinq étapes narratives, ce sont la situation initiale, l’action se déclenche, l’action se développe, l’action se dénoue et la situation finale. L’histoire du roman se termine par la fin heureuse. Le personnage principal du roman est Blériot. Les personnages supplémentaires du roman sont Nora, Murphy et Sabine. L’histoire a eu lieu à Paris et à Londres. Il se déroule depuis l’appel de Nora à Blériot dans le jour de l’ascension jusqu’au jour où Nora est venu à l’appartement de Blériot à Nice. Les cadres sociaux du roman sont la classe sociale du personnage principale qui est dans la classe inférieur et les conditions sociales de la société européenne moderne, en particulier la France et la Grande-Bretagne. (2) les éléments intrinsèques de ce roman sont relier et liée par le thème forme une histoire cohérente. Le thème majeur du roman est le désire de l'amour qui est sans fin, tandis que les thèmes mineurs sont l'infidélité, la trahison, et la liberté. (3) basé sur l'analyse sémiotique du roman, on trouve que le désir sans fin de l’homme fait la vie dans le monde se sent très brièvement pour le satisfaire, et il doit y avoir une lutte et un sacrifice pour atteindre des désirs qui créent le bonheur dans la vie

Cost effectiveness of a community based prevention and treatment of acute malnutrition programme in Mumbai slums, India

Children in slums are at high risk of acute malnutrition and death. Cost-effectiveness of community-based management of severe acute malnutrition programmes has been demonstrated previously, but there is limited evidence in the context of urban slums where programme cost structure is likely to vary tremendously. This study assessed the cost-utility of adding a community based prevention and treatment for acute malnutrition intervention to Government of India Integrated Child Development Services (ICDS) standard care for children in Mumbai slums. The intervention is delivered by community health workers in collaboration with ICDS Anganwadi community health workers. The analysis used a decision tree model to compare the costs and effects of the two options: standard ICDS services with the intervention and prevention versus standard ICDS services alone. The model used outcome and cost data from the Society for Nutrition, Education & Health Action's Child Health and Nutrition programme in Mumbai slums, which delivered services to 12,362 children over one year from 2013 to 2014. An activity-based cost model was used, with calculated costs based on programme financial records and key informant interviews. Cost data were coupled with programme effectiveness data to estimate disability adjusted life years (DALYs) averted. The community based prevention and treatment programme averted 15,016 DALYs (95% Uncertainty Interval [UI]: 12,246-17,843) at an estimated cost of $23 per DALY averted (95%UI:19-28) and was thus highly cost-effective. This study shows that ICDS Anganwadi community health workers can work efficiently with community health workers to increase the prevention and treatment coverage in slums in India and can lead to policy recommendations at the state, and potentially the national level, to promote such programmes in Indian slums as a cost-effective approach to tackling moderate and severe acute malnutrition

Cost effectiveness of a community based prevention and treatment of acute malnutrition programme in Mumbai slums, India

Children in slums are at high risk of acute malnutrition and death. Cost-effectiveness of community-based management of severe acute malnutrition programmes has been demonstrated previously, but there is limited evidence in the context of urban slums where programme cost structure is likely to vary tremendously. This study assessed the cost-utility of adding a community based prevention and treatment for acute malnutrition intervention to Government of India Integrated Child Development Services (ICDS) standard care for children in Mumbai slums. The intervention is delivered by community health workers in collaboration with ICDS Anganwadi community health workers. The analysis used a decision tree model to compare the costs and effects of the two options: standard ICDS services with the intervention and prevention versus standard ICDS services alone. The model used outcome and cost data from the Society for Nutrition, Education & Health Action’s Child Health and Nutrition programme in Mumbai slums, which delivered services to 12,362 children over one year from 2013 to 2014. An activity-based cost model was used, with calculated costs based on programme financial records and key informant interviews. Cost data were coupled with programme effectiveness data to estimate disability adjusted life years (DALYs) averted. The community based prevention and treatment programme averted 15,016 DALYs (95% Uncertainty Interval [UI]: 12,246–17,843) at an estimated cost of $23 per DALY averted (95%UI:19–28) and was thus highly cost-effective. This study shows that ICDS Anganwadi community health workers can work efficiently with community health workers to increase the prevention and treatment coverage in slums in India and can lead to policy recommendations at the state, and potentially the national level, to promote such programmes in Indian slums as a cost-effective approach to tackling moderate and severe acute malnutrition

Cost effectiveness of a community based prevention and treatment of acute malnutrition programme in Mumbai slums, India

Children in slums are at high risk of acute malnutrition and death. Cost-effectiveness of community-based management of severe acute malnutrition programmes has been demonstrated previously, but there is limited evidence in the context of urban slums where programme cost structure is likely to vary tremendously. This study assessed the cost-utility of adding a community based prevention and treatment for acute malnutrition intervention to Government of India Integrated Child Development Services (ICDS) standard care for children in Mumbai slums. The intervention is delivered by community health workers in collaboration with ICDS Anganwadi community health workers. The analysis used a decision tree model to compare the costs and effects of the two options: standard ICDS services with the intervention and prevention versus standard ICDS services alone. The model used outcome and cost data from the Society for Nutrition, Education & Health Action's Child Health and Nutrition programme in Mumbai slums, which delivered services to 12,362 children over one year from 2013 to 2014. An activity-based cost model was used, with calculated costs based on programme financial records and key informant interviews. Cost data were coupled with programme effectiveness data to estimate disability adjusted life years (DALYs) averted. The community based prevention and treatment programme averted 15,016 DALYs (95% Uncertainty Interval [UI]: 12,246-17,843) at an estimated cost of $23 per DALY averted (95%UI:19-28) and was thus highly cost-effective. This study shows that ICDS Anganwadi community health workers can work efficiently with community health workers to increase the prevention and treatment coverage in slums in India and can lead to policy recommendations at the state, and potentially the national level, to promote such programmes in Indian slums as a cost-effective approach to tackling moderate and severe acute malnutrition

Human cardiac tissue in a microperfusion chamber simulating extracorporeal circulation - ischemia and apoptosis studies

<p>Abstract</p> <p>Background</p> <p>After coronary artery bypass grafting ischemia/reperfusion injury inducing cardiomyocyte apoptosis may occur. This surgery-related inflammatory reaction appears to be of extreme complexity with regard to its molecular, cellular and tissue mechanisms and many studies have been performed on animal models. However, finding retrieved from animal studies were only partially confirmed in humans. To investigate this phenomenon and to evaluate possible therapies in vitro, adequate human cardiomyocyte models are required. We established a tissue model of human cardiomyocytes preserving the complex tissue environment. To our knowledge human cardiac tissue has not been investigated in an experimental setup mimicking extracorporeal circulation just in accordance to clinical routine, yet.</p> <p>Methods</p> <p>Cardiac biopsies were retrieved from the right auricle of patients undergoing elective coronary artery bypass grafting before cardiopulmonary bypass. The extracorporeal circulation was simulated by submitting the biopsies to varied conditions simulating cardioplegia (cp) and reperfusion (rep) in a microperfusion chamber. Cp/rep time sets were 20/7, 40/13 and 60/20 min. For analyses of the calcium homoeostasis the fluorescent calcium ion indicator FURA-2 and for apoptosis detection PARP-1 cleavage immunostaining were employed. Further the anti-apoptotic effect of carvedilol [10 μM] was investigated by adding into the perfusate.</p> <p>Results</p> <p>Viable cardiomyocytes presented an intact calcium homoeostasis under physiologic conditions. Following cardioplegia and reperfusion a time-dependent elevation of cytosolic calcium as a sign of disarrangement of the calcium homoeostasis occurred. PARP-1 cleavage also showed a time-dependence whereas reperfusion had the highest impact on apoptosis. Cardioplegia and carvedilol could reduce apoptosis significantly, lowering it between 60-70% (p < 0.05).</p> <p>Conclusions</p> <p>Our human cardiac preparation served as a reliable cellular model tool to study apoptosis in vitro. Decisively cardiac tissue from the right auricle can be easily obtained at nearly every cardiac operation avoiding biopsying of the myocardium or even experiments on animals.</p> <p>The apoptotic damage induced by the ischemia/reperfusion stimulus could be significantly reduced by the cold crystalloid cardioplegia. The additional treatment of cardiomyocytes with a non-selective β-blocker, carvedilol had even a significantly higher reduction of apoptotis.</p