DISCUSSION ON DESIGNING AND ASSESSING DISASTER PREVENTION EDUCATION BASED ON ID FOCUSING ON THE LEADING EVACUEES DRILLS USING RED/BLUE PAPERPERBOARD

本研究では筆者らが行った「青赤紙を用いた率先避難訓練」の防災教育を事例に，よりよい防災教育へ改善していくための問題点と，その改善策を検討した．その結果，本事例の問題点として(1)率先避難者の役割をどのように教えるのかといった「教授方略」に注力し，「目標」や「評価」を決めるための学習者分析を十分に行わなかったこと，(2)「目標」や「評価」をあいまいなままに教育を行ったことで，学習者が理解したことと教育者が教えたいことにズレがあった可能性を見出すことができた．インストラクショナルデザイン（以下ID）を用いることでこの問題点の改善策を提示し，「目標」⇒「評価」⇒「教授方略」の順番で教育を設計することが，よりよい防災教育へ改善していくために重要であることを，防災教育の具体的事例に基づき示すことができた．Based on the example of a disaster preparedness education program implemented by the authors, leading evacuees drills using red/blue paperboard, this study examined the problems of the program and their remedial measures for improving disaster preparedness education. As a result, the following two problems were identified: 1) the focus was on the teaching strategies, such as methods for teaching the roles of leading evacuees, and learner analysis for setting the goals and the assessment criteria was not fully conducted, 2) there might have been a gap between what the instructors wanted to teach and what the learners understood due to the fact that the program was implemented without setting clear goals and assessment criteria. Based on a specific example of disaster preparedness education, it was demonstrated that in order to improve disaster preparedness education, it is important to present remedial measures for the problems of the program using instructional design (ID) and to design the education program in the order of goals, assessment criteria, and finally teaching strategies

Uji Aktivitas Antibakteri dari Ekstrak n-Heksana dan Kloroform Daun Sirsak (Annona muricate L.) Terhadap Pertumbuhan Bakteri Staphylococcus aureus Secara In Vitro

Penelitian ini bertujuan untuk mengetahui aktivitas antibakteri daun sirsak terhadap Staphylococcus aureus. Ekstraksi diproses dengan metode maserasi terlebih dahulu dengan n-heksana kemudian kloroform. Aktivitas antibakteri diukur secara in vitro dengan menggunakan metode difusi agar menggunakan paperdisk. Aktivitas antibakteri diuji dengan analisis variansi (ANOVA) dengan nilai signifikansi 0,5% untuk mengetahui pengobatan mana yang memiliki efek atau berbeda secara signifikan dengan dosis masing-masing 300 mg/ml, 250 mg/ml, 200 mg/ml, 150 mg/ml, 100 mg/ml dan menghasilkan aktivitas antibakteri dengan zona hambat berturut-turut 16,70 mm; 14,05 mm; 11,45 mm; 9,85 mm; 3,00. Hasil pemeriksaan aktivitas antibakteri menunjukkan ekstrak n-heksana dan kloroform dapat menghambat pertumbuhan Staphylococcus aureus pada konsentrasi 250 mg / ml dengan diameter zona hambat 14,05 mm. Ekstrak dengan aktivitas tertinggi ditentukan konsentrasi hambat tumbuh minimum (MIC). Ekstrak kloroform daun sirsak dari bakteri Staphylococcus aureus yang berada pada konsentrasi 100 mg/ml dengan zona hambat 3,00 mm

Uji Aktivitas Antibakteri dari Ekstrak N-Heksana dan Kloroform Daun Sirsak (annona muricate I) Terhadap Pertumbuhan Bakteri Staphylococcus aureus Secara In Vitro

Penelitian ini bertujuan untuk mengetahui aktivitas antibakteri daun sirsak terhadap Staphylococcus aureus. Ekstraksi diproses dengan metode maserasi terlebih dahulu dengan n-heksana kemudian kloroform. Aktivitas antibakteri diukur secara in vitro dengan menggunakan metode difusi agar menggunakan paperdisk. Aktivitas antibakteri diuji dengan analisis variansi (ANOVA) dengan nilai signifikansi 0,5% untuk mengetahui pengobatan mana yang memiliki efek atau berbeda secara signifikan dengan dosis masing-masing 300 mg/ml, 250 mg/ml, 200 mg/ml, 150 mg/ml, 100 mg/ml dan menghasilkan aktivitas antibakteri dengan zona hambat berturut-turut 16,70 mm; 14,05 mm; 11,45 mm; 9,85 mm; 3,00. Hasil pemeriksaan aktivitas antibakteri menunjukkan ekstrak n-heksana dan kloroform dapat menghambat pertumbuhan Staphylococcus aureus pada konsentrasi 250 mg / ml dengan diameter zona hambat 14,05 mm. Ekstrak dengan aktivitas tertinggi ditentukan konsentrasi hambat tumbuh minimum (MIC). Ekstrak kloroform daun sirsak dari bakteri Staphylococcus aureus yang berada pada konsentrasi 100 mg/ml dengan zona hambat 3,00 mm. Keywords : Annona muricata L; antibakteri; ekstrak kloroform; Staphylococcus aureu

Functional Analysis of the Leading Malaria Vaccine Candidate AMA-1 Reveals an Essential Role for the Cytoplasmic Domain in the Invasion Process

A key process in the lifecycle of the malaria parasite Plasmodium falciparum is the fast invasion of human erythrocytes. Entry into the host cell requires the apical membrane antigen 1 (AMA-1), a type I transmembrane protein located in the micronemes of the merozoite. Although AMA-1 is evolving into the leading blood-stage malaria vaccine candidate, its precise role in invasion is still unclear. We investigate AMA-1 function using live video microscopy in the absence and presence of an AMA-1 inhibitory peptide. This data reveals a crucial function of AMA-1 during the primary contact period upstream of the entry process at around the time of moving junction formation. We generate a Plasmodium falciparum cell line that expresses a functional GFP-tagged AMA-1. This allows the visualization of the dynamics of AMA-1 in live parasites. We functionally validate the ectopically expressed AMA-1 by establishing a complementation assay based on strain-specific inhibition. This method provides the basis for the functional analysis of essential genes that are refractory to any genetic manipulation. Using the complementation assay, we show that the cytoplasmic domain of AMA-1 is not required for correct trafficking and surface translocation but is essential for AMA-1 function. Although this function can be mimicked by the highly conserved cytoplasmic domains of P. vivax and P. berghei, the exchange with the heterologous domain of the microneme protein EBA-175 or the rhoptry protein Rh2b leads to a loss of function. We identify several residues in the cytoplasmic tail that are essential for AMA-1 function. We validate this data using additional transgenic parasite lines expressing AMA-1 mutants with TY1 epitopes. We show that the cytoplasmic domain of AMA-1 is phosphorylated. Mutational analysis suggests an important role for the phosphorylation in the invasion process, which might translate into novel therapeutic strategies

The Essentials of Protein Import in the Degenerate Mitochondrion of Entamoeba histolytica

Several essential biochemical processes are situated in mitochondria. The metabolic transformation of mitochondria in distinct lineages of eukaryotes created proteomes ranging from thousands of proteins to what appear to be a much simpler scenario. In the case of Entamoeba histolytica, tiny mitochondria known as mitosomes have undergone extreme reduction. Only recently a single complete metabolic pathway of sulfate activation has been identified in these organelles. The E. histolytica mitosomes do not produce ATP needed for the sulfate activation pathway and for three molecular chaperones, Cpn60, Cpn10 and mtHsp70. The already characterized ADP/ATP carrier would thus be essential to provide cytosolic ATP for these processes, but how the equilibrium of inorganic phosphate could be maintained was unknown. Finally, how the mitosomal proteins are translocated to the mitosomes had remained unclear. We used a hidden Markov model (HMM) based search of the E. histolytica genome sequence to discover candidate (i) mitosomal phosphate carrier complementing the activity of the ADP/ATP carrier and (ii) membrane-located components of the protein import machinery that includes the outer membrane translocation channel Tom40 and membrane assembly protein Sam50. Using in vitro and in vivo systems we show that E. histolytica contains a minimalist set up of the core import components in order to accommodate a handful of mitosomal proteins. The anaerobic and parasitic lifestyle of E. histolytica has produced one of the simplest known mitochondrial compartments of all eukaryotes. Comparisons with mitochondria of another amoeba, Dictystelium discoideum, emphasize just how dramatic the reduction of the protein import apparatus was after the loss of archetypal mitochondrial functions in the mitosomes of E. histolytica

Insufficient sensitivity of laser desorption-time of flight mass spectrometry-based detection of hemozoin for malaria screening

Laser desorption-time of flight (LD-TOF) mass spectrometry-based detection of hemozoin was assessed for its performance characteristics as a rapid screening test for malaria. In spite of good specificity of >95%, poor sensitivity of 80.2% for microscopically positive samples makes the easy-to-apply and rapid approach unsuitable for the routine diagnostic setting

Pellicle formation in the malaria parasite

The intraerythrocytic developmental cycle of Plasmodium falciparum is completed with the release of up to 32 invasive daughter cells, the merozoites, into the blood stream. Before release, the final step of merozoite development is the assembly of the cortical pellicle, a multi-layered membrane structure. This unique apicomplexan feature includes the inner membrane complex (IMC) and the parasite's plasma membrane. A dynamic ring structure, referred to as the basal complex, is part of the IMC and helps to divide organelles and abscises in the maturing daughter cells. Here, we analyze the dynamics of the basal complex of P. falciparum. We report on a novel transmembrane protein of the basal complex termed BTP1, which is specific to the genus Plasmodium. It colocalizes with the known basal complex marker protein MORN1 and shows distinct dynamics as well as localization when compared to other IMC proteins during schizogony. Using a parasite plasma membrane marker cell line, we correlate dynamics of the basal complex with the acquisition of the maternal membrane. We show that plasma membrane invagination and IMC propagation are interlinked during the final steps of cell division

Transcriptional profiling of growth perturbations of the human malaria parasite Plasmodium falciparum

Functions have yet to be defined for the majority of genes of Plasmodium falciparum, the agent responsible for the most serious form of human malaria. Here we report changes in P. falciparum gene expression induced by 20 compounds that inhibit growth of the schizont stage of the intraerythrocytic development cycle. In contrast with previous studies, which reported only minimal changes in response to chemically induced perturbations of P. falciparum growth, we find that ~59% of its coding genes display over three-fold changes in expression in response to at least one of the chemicals we tested. We use this compendium for guilt-by-association prediction of protein function using an interaction network constructed from gene co-expression, sequence homology, domain-domain and yeast two-hybrid data. The subcellular localizations of 31 of 42 proteins linked with merozoite invasion is consistent with their role in this process, a key target for malaria control. Our network may facilitate identification of novel antimalarial drugs and vaccines.<br /

Ancient human sialic acid variant restricts an emerging zoonotic malaria parasite

Plasmodium knowlesi is a zoonotic parasite transmitted from macaques causing malaria in humans in Southeast Asia. Plasmodium parasites bind to red blood cell (RBC) surface receptors, many of which are sialylated. While macaques synthesize the sialic acid variant N-glycolylneuraminic acid (Neu5Gc), humans cannot because of a mutation in the enzyme CMAH that converts N-acetylneuraminic acid (Neu5Ac) to Neu5Gc. Here we reconstitute CMAH in human RBCs for the reintroduction of Neu5Gc, which results in enhancement of P. knowlesi invasion. We show that two P. knowlesi invasion ligands, PkDBPβ and PkDBPγ, bind specifically to Neu5Gc-containing receptors. A human-adapted P. knowlesi line invades human RBCs independently of Neu5Gc, with duplication of the sialic acid-independent invasion ligand, PkDBPα and loss of PkDBPγ. Our results suggest that absence of Neu5Gc on human RBCs limits P. knowlesi invasion, but that parasites may evolve to invade human RBCs through the use of sialic acid-independent pathways