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Abstrak :
[ABSTRAK
Pendahuluan: Infeksi dengue merupakan salah satu penyakit endemik di daerah tropis dan subtropis yang disebabkan oleh virus dengue (DENV). Hingga saat ini belum ada antiviral yang efektif untuk infeksi dengue. Penyebaran dan sirkulasi serotipe DENV berfariasi di setiap lokasi geografi, hal ini menyulitkan dalam melakukan evaluasi vaksin DENV. Oleh karena itu perlu dikembangkan kandidat vaksin DENV menggunakan strain Indonesia supaya dapat memberikan proteksi maksimal. Pada peneltian ini dikembangkan kandidat vaksin DNA tetravalen DENV berbasis gen prM-E DENV strain Indonesia. Metode: Konstruksi plasmid rekombinan kandidat vaksi dilakukan dengan cara menyisipkan gen prM-E setiap serotipe DENV ke dalam vektor pUMVC4a. Gen prM-E DENV merupakan strain Indonesia, yang diamplifikasi dari serum pasien yang terinfeksi dengan virus ini. Kemampuan plasmid rekombinan mengekspresikan protein prM-E DENV diuji di sel mamalia. Kemampuan kandidat vaksin menginduksi respon imun humoral dievaluasi secara monovalen dan tetravalen di mencit jenis ddY. Titer IgG anti dengue diperiksa menggunakan teknik ELISA, sedangkan titer antibodi netralisasi di tentukan dengan uji FRNT. Proteksi vaksin terhadap mencit yang diimunisasi dievaluasi dengan melakukan uji tantang menggunakan sel K562 yang diinfeksi DENV-2. Viremi virus di tentukan dengan menggunakan teknik foccus assay. Hasil: Konstruksi plasmid rekombinan kandidat vaksin DENV-1 dan DENV-3 sudah berhasil dilakukan. Plasmid dapat mengekspresikan protein prM-E DENV di sel mamalia, namun karakteristik dan kinetik protein masih belum dapat diketahui dengan jelas. Keempat kandidat vaksin DNA yang sedang dikembangkan dapat menginduksi respon imun, baik secara monovalen maupun tetravalen. Imunisasi secara tetravalen dapat memberikan proteksi pada mencit yang diuji tantang dengan sel K562 yang diinfeksi dengan DENV-2.;

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ABSTRACT
Introduction: Dengue infections are caused by dengue viruses (DENV) and are endemic in tropical and subtropical regions. At present, there is no effective antiviral treatment for dengue infection. Distribution and circulation of DENV serotypes varies by geographic location, it is difficult to evaluate DENV vaccine. Therefore, it is necessary to develop a vaccine candidate DENV using Indonesian strains in order to provide maximum protection. However, in this study, we constructed a recombinant plasmid-based prM-E gene from the Indonesia strain as a DENV DNA vaccine candidate. Methode: The recombinant plasmid was prepared by inserting the prM-E gene from each DENV serotypes into the plasmid backbone pUMVC4a. prM-E gene an Indonesia strain, which was amplified from patient sera infected with DENV. The ability of the recombinant plasmid expressing the prM-E DENV protein tested in mammalian cells. The ability of candidate vaccines induce humoral immune responses were evaluated monovalent and tetravalent in ddY mice. IgG titers of anti-dengue examined using ELISA technique, while neutralizing antibody titers determined with FRNT test. Vaccine protection against the immunized mice was evaluated by conducting challenge test using K562 cells infected by DENV-2. Viremia was determined by using the foccus assay. Result: Construction of recombinant plasmid vaccine candidate DENV-1 and DENV-3 was successfully performed. Plasmids can express prM-E DENV proteins in mammalian cells, but the characteristics and kinetics of protein still can not clearly known. Fourth DNA vaccine candidate that is being developed to induce an immune response, either monovalent or tetravalent. Tetravalent immunization may provide protection in mice challenged tested with K562 cells infected with DENV-2.;Introduction: Dengue infections are caused by dengue viruses (DENV) and are endemic in tropical and subtropical regions. At present, there is no effective antiviral treatment for dengue infection. Distribution and circulation of DENV serotypes varies by geographic location, it is difficult to evaluate DENV vaccine. Therefore, it is necessary to develop a vaccine candidate DENV using Indonesian strains in order to provide maximum protection. However, in this study, we constructed a recombinant plasmid-based prM-E gene from the Indonesia strain as a DENV DNA vaccine candidate. Methode: The recombinant plasmid was prepared by inserting the prM-E gene from each DENV serotypes into the plasmid backbone pUMVC4a. prM-E gene an Indonesia strain, which was amplified from patient sera infected with DENV. The ability of the recombinant plasmid expressing the prM-E DENV protein tested in mammalian cells. The ability of candidate vaccines induce humoral immune responses were evaluated monovalent and tetravalent in ddY mice. IgG titers of anti-dengue examined using ELISA technique, while neutralizing antibody titers determined with FRNT test. Vaccine protection against the immunized mice was evaluated by conducting challenge test using K562 cells infected by DENV-2. Viremia was determined by using the foccus assay. Result: Construction of recombinant plasmid vaccine candidate DENV-1 and DENV-3 was successfully performed. Plasmids can express prM-E DENV proteins in mammalian cells, but the characteristics and kinetics of protein still can not clearly known. Fourth DNA vaccine candidate that is being developed to induce an immune response, either monovalent or tetravalent. Tetravalent immunization may provide protection in mice challenged tested with K562 cells infected with DENV-2., Introduction: Dengue infections are caused by dengue viruses (DENV) and are endemic in tropical and subtropical regions. At present, there is no effective antiviral treatment for dengue infection. Distribution and circulation of DENV serotypes varies by geographic location, it is difficult to evaluate DENV vaccine. Therefore, it is necessary to develop a vaccine candidate DENV using Indonesian strains in order to provide maximum protection. However, in this study, we constructed a recombinant plasmid-based prM-E gene from the Indonesia strain as a DENV DNA vaccine candidate. Methode: The recombinant plasmid was prepared by inserting the prM-E gene from each DENV serotypes into the plasmid backbone pUMVC4a. prM-E gene an Indonesia strain, which was amplified from patient sera infected with DENV. The ability of the recombinant plasmid expressing the prM-E DENV protein tested in mammalian cells. The ability of candidate vaccines induce humoral immune responses were evaluated monovalent and tetravalent in ddY mice. IgG titers of anti-dengue examined using ELISA technique, while neutralizing antibody titers determined with FRNT test. Vaccine protection against the immunized mice was evaluated by conducting challenge test using K562 cells infected by DENV-2. Viremia was determined by using the foccus assay. Result: Construction of recombinant plasmid vaccine candidate DENV-1 and DENV-3 was successfully performed. Plasmids can express prM-E DENV proteins in mammalian cells, but the characteristics and kinetics of protein still can not clearly known. Fourth DNA vaccine candidate that is being developed to induce an immune response, either monovalent or tetravalent. Tetravalent immunization may provide protection in mice challenged tested with K562 cells infected with DENV-2.]

Abstrak :
Tujuan: Metabolit yang dihasilkan oleh mikroorganisme merupakan sumber yang potensial untuk dieksplorasi guna memperoleh senyawa aktif antimikroba, salah satunya adalah kelompok biosurfaktan lipopeptida. Berbagai senyawa lipopeptida mempunyai aktifitas biologi yang tinggi. seperti aktifitas antikanker, antivirus, antipembekuan darah, immunomodulator, antiadesif, antiparasit, antibakteri dan antijamur. Tujuan dari penelitian ini adalah untuk mengeksplorasi kekayaan biodiversitas nasional dengan melakukan isolasi dan skrining mikroba penghasil biosurfaktan lipopeptida serta karakterisasi dari lipopeptida yang dihasilkan sebagai antimikroba untuk aplikasi di bidang biomedis. Metode: Isolasi dan skrining mikroba penghasil biosurfaktan lipopeptida dilakukan dengan mengambil sampel berupa tanah dan air dari lokasi yang tercemar minyak baik di darat maupun di laut. Isolat terpilih digunakan dalam proses fermentasi untuk produksi lipopeptida. Uji aktifitas antibakteri dilakukan terhadap beberapa bakteri uji gram positif maupun negatif. Senyawa aktif lipopeptida yang dihasilkan diisolasi dan dikarakterisasi strukturnya menggunakan spektrometri massa. Optimasi produksi juga dilakukan guna memperoleh kondisi proses yang optimal menggunakan Respon Surface Methodology. Studi efek kombinasi senyawa lipopeptida dengan antibiotik lain dilakukan menggunakan metode double disk dan metode checkerboard. Hasil: Diperoleh mikroba penghasil biosurfaktan lipopeptida Bacillus amyloliquefaciens MD4-12 yang diisolasi dari lokasi di sekitar kilang minyak Pertamina di Palembang. Lipopeptida yang dihasilkan mampu menghambat pertumbuhan bakteri uji gram positif dan negatif secara in-vitro menggunakan metode difusi cakram termasuk bakteri resisten MRSA (methicillin resistant strain Staphylococcus aureus) dan E. coli ATCC 32518, bakteri penghasil betalaktamase. Hasil karakterisasi senyawa menunjukkan bahwa lipopeptida yang dihasilkan adalah surfaktin homolog yang terdiri dari C12-C17 surfaktin. Pada optimasi produksi senyawa surfaktin diperoleh peningkatan produksi sebesar 2,4 kali dari perolehan sebelum dilakukan optimasi, yaitu dari 0,51 g/L menjadi 1,21 g/L. Studi kombinasi senyawa lipopeptida dengan antibiotik ampisilin menunjukkan efek sinergi dan aditif dalam menghambat pertumbuhan bakteri uji Pseudomonas aeruginosa ATCC 27853. Nilai FIC indeks yang diperoleh berkisar antara 0,31 ? 0,63. Penelitian ini menunjukkan potensi senyawa lipopeptida surfaktin sebagai antibakteri untuk aplikasi di bidang biomedis.

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Objective. Metabolites produced by microorganisms are potential sources to be explored to obtain biological active compound. One of them belongs to lipopeptide biosurfactants group. Various active compounds of lipopeptide have high biological activity for biomedical application such as anticancer, antiviral, inhibit fibrin clot formation, immunomodulators, antiadesif, antiparasitic, antibacterial and antifungi. The aim of this study was to explore the national biodiversity to obtained microbial lipopeptide biosurfactants producers with antimicrobial activity. Methods: To isolate and screen lipopeptide-producing microbes, soil and water samples were taken from oil contaminated from terrestrial and marine. Selected isolates were used for fermentation to produce lipopeptides. Active compound were isolated and characterized structurally by mass spectrometry. Optimization of production was also carried out to obtain optimal process conditions using the Response Surface Methodology. The effect of lipopeptides combination with other antibiotics for antimicrobial activity were performed against several test bacteria using double disc and checkerboard methods. Result: Lipopeptide biosurfactant-producing microbe was obtained from the soil sample around Pertamina oil refinery plant at Palembang. Furthermore the isolate was identified as Bacillus amyloliquefaciens MD4-12. The Lipopeptides capable to inhibit the growth of gram positive and negative tests bacteria in-vitro, including resistant bacteria MRSA (methicillin resistant strain Staphylococcus aureus) and E. coli ATCC 32518, a betalactamase-producing strain, using diffusion disc method. Characterization of lipopeptide compounds using mass spectrometry showed that the lipopeptide is surfactin homolog consist of C12-C17 surfactin. Optimum medium composition was obtained during optimization of surfactin production using respon surface methodology. Surfactin production increased 2.4 times from 0.51 g/L, prior to optimization, to 1.21 g/L. Combination lipopeptide with ampicillin for antibacterial activity showed synergistic and additive effects against the test bacteria Pseudomonas aeruginosa ATCC 27853. The range of FIC index is 0.31 to 0.63. This research showed that surfactin lipopeptide have great potency for antimicrobial activity for biomedical application.

Abstrak :
COVID-19 merupakan penyakit penyebab pandemi pada akhir 2019. Perbedaan manifestasi klinis pada infeksi SARS-CoV-2 ini memicu banyak pertanyaan di kalangan peneliti dan medis. Perbedaan klinis COVID-19 tersebut dapat dipicu oleh faktor hospes, patogen maupun lingkungan. Infeksi SARS-CoV-2 terutama melalui saluran napas atas, tempat kolonisasi mikroba komensal dan patogen. Bagaimana interaksi antara mikroba yang berkolonisasi dengan SARS-CoV-2 dalam menimbulkan respons inflamasi di saluran napas atas masih belum diketahui dengan jelas. Penelitian ini bertujuan menganalisis hubungan antara karakteristik mikrobiota, serta rasio kadar sitokin pro- dan anti-inflamasi dari saluran napas atas dengan beratnya COVID-19. Penelitian ini merupakan studi potong lintang menggunakan 74 swab nasofaring dan orofaring di dalam viral transport medium (VTM) dari pasien COVID-19 berusia 18–64 tahun. Profil mikrobiota di saluran napas atas dan kadar IL-6, IL-1β, IFN-γ, TNF-α dan IL-10 diperiksa dengan metode sekuensing 16S ribosomal RNA dan Luminex assay, secara berurutan. Selanjutnya dilakukan analisis hubungan antara beratnya COVID-19 dengan OTU, keragaman alfa dan beta dari mikrobiota saluran napas atas. Lima filum terbanyak di saluran napas pasien COVID-19 di Indonesia berusia 18-64 tahun adalah Firmicutes (32,3%), Bacteroidota (27,1%), Fusobacteriota (15,2%), Proteobacteria (15,1%) dan Actinobacteria (7,1%). Analisis indeks Shannon dan ACE menunjukkan bahwa tidak ada penurunan keragaman microbiota saluran napas atas dengan bertambah beratnya penyakit. Namun, ada perbedaan bermakna keragaman beta pada mikrobiota saluran napas atas antara COVID-19 ringan dan berat. Keberlimpahan filum Firmicutes (p = 0,012), dan genus Streptococcus (p = 0,033) dan Enterococcus (p = 0,031) lebih tinggi pada COVID-19 berat dibandingkan yang ringan, sedangkan keberlimpahan filum Fusobacteriota (p = 0,021), Proteobacteria (p = 0,030), Campilobacterota (p = 0,027), genus Neisseria (p = 0,008), dan Fusobacterium (p = 0,064), spesies Porphyromonas gingivalis (p = 0,018), Fusobacterium periodonticum (p = 0,001) dan Fusobacterium nucleatum (p = 0,022) lebih tinggi pada COVID-19 ringan dibandingkan berat. Keberadaan bakteri Prevotella buccae (p = 0,005) dan Prevotella disiens (p = 0,043) lebih rendah pada COVID-19 berat. Rasio TNF-α/IL-10 lebih tinggi pada COVID-19 berat (p < 0.05). Selanjutnya, rasio IL-6/IL-10, IFN-γ/IL-10, dan IL-1β/IL-10 juga lebih tinggi pada COVID-19 berat, namun tidak berbeda bermakna jika dikaitkan dengan beratnya penyakit. Penelitian ini mendukung adanya hubungan antara karakteristik mikrobiota di saluran napas atas dengan beratnya COVID-19 pada pasien dewasa. Studi lebih lanjut diperlukan untuk memeriksa mekanisme bagaimana mikrobiota mencegah beratnya COVID-19. Rasio TNF-α/IL-10 dari saluran napas dapat menjadi prediktor beratnya penyakit dan sebagai alternatif pemeriksaan kadar sitokin pada COVID-19 yang kurang invasif dibandingkan serum. ......COVID-19 is a disease that caused a pandemic at the end of 2019. Clinical manifestations difference in SARS-CoV-2 infection has raised many questions in research and medical provider. The clinical differences in COVID-19 can be triggered by host, pathogen and environmental factors. SARS-CoV-2 mainly enters through the upper airway, with colonization of commensal and pathogenic microbes. How the interaction between colonized microbes and SARS-CoV-2 in causing an inflammatory response in the upper airway is still not clearly known. Therefore, we examined the association between the diversity of microbiota, pro- and anti-inflammatory cytokines ratio of upper respiratory and COVID-19 severity. This research is an observational cross-sectional study using 74 nasopharyngeal and oropharyngeal swabs in viral transport medium from COVID-19 patients aged 18-64 years. We examined microbiota profile in the upper airway using 16S ribosomal RNA sequencing method and levels of IL-6, IL-1β, IFN-γ, TNF-α and IL-10 were examined by Luminex assay. We also examined the association between COVID-19 severities with OTU analysis, alpha and beta diversity of upper respiratory microbiota. The top five phyla in upper respiratory tract of Indonesian COVID-19 patients with aged of 18–64 years old were Firmicutes (32,3%), Bacteroidota (27,1%), Fusobacteriota (15,2%), Proteobacteria (15,1%) and Actinobacteria (7,1%). Shannon and ACE index analysis showed no decline of microbiota diversity in upper airway with the increase of disease severity. However, there were significant differences of beta diversity in the upper airway microbiota between mild and severe COVID-19. The abundance of the Firmicutes phylum (p = 0,012), Streptococcus (p = 0,033) and Enterococcus (p = 0,031) genera were significantly higher in severe COVID-19 than mild, while the abundance of the Fusobacteriota (p = 0,021), Proteobacteria (p=0,030), and Campilobacterota (p = 0,027) phyla, Neisseria (p = 0,008), and Fusobacterium (p = 0,064) genera, Porphyromonas gingivalis (p = 0,018), Fusobacterium periodonticum (p = 0,001) and Fusobacterium nucleatum (p = 0,022) species were significantly higher in mild. The presence of Prevotella buccae (p=0.005) and Prevotella disiens (p=0.043) bacteria was lower in severe COVID-19. The TNF-α/IL-10 ratio was significantly higher in severe COVID-19 (p < 0.05). Furthermore, IL-6/IL-10, IFN-γ/IL-10, and IL-1β/IL-10 ratio was also higher in severe, but those were not significantly related to the disease severity. This research supports the relationship between the severity of COVID-19 and microbiota diversity in the upper airway in adults. Further studies are needed to examine the mechanism by which microbiota prevents the COVID-19 severities. The ratio of TNF-α/IL-10 from upper airway swab may be as a predictor of disease severity and alternative for examining cytokine levels in COVID-19 which is less invasive than serum.