Induced-Coagulated Plasma-Fibrin Gels as a Biological Scaffold for Cell Attachment and Proliferation of Umbilical Cord-Derived Mesenchymal Stem Cells (UC-MSC)

Fibrin gels are an ideal natural biological scaffold for tissue engineering because they are biocompatible,biodegradable, and have many biological surface markers. However, most research on fi brin gels used commercialfi brin kits that could be costly and limited in some areas. In this study, fi brin gels were made by inducing bloodcoagulation by adding a common diagnostic kit to assess the time for blood to clot, called activated partialthromboplastin time (aPTT). This induced coagulated plasma (iCoplas)-fi brin gels was evaluated for its ability toenhance biological activity of umbilical cord-derived mesenchymal stem cell (UC-MSC), which were cell attachmentand proliferation. Fibrinogen concentration had infl uence on cell attachment, where only 50% of the cells couldattach to 77 mg/dl fi brinogen gels whereas 93% cells adhered to 154 mg/dl fi brin gels. There were no signifi cantdifferences in cell proliferation on polysterene culture dish and fi brin gels (p>0.05). These results showed thatiCoplas-fi brin gels could be used as a fi brin-based scaffold, yielding no signifi cant difference than polysterene-tissueculture dish cultures in cell attachment and cell proliferation on 154 mg/dl fi brinogen concentration

Kajian aspek keamanan nyamuk Aedes aegypti Linnaeus ber-Wolbachia di Yogyakarta, Indonesia

Dengue prevention efforts are limited to the control strategies of its vector and the management of breeding sites. New alternatives for dengue vector control that are sustainable and more environmentally friendly are needed to complement the government’s current efforts. Research on Wolbachia-infected Aedes aegypti Linnaeus mosquitoes as an alternative biocontrol strategy has been performed in Yogyakarta City. However, one of the concerns of the community members and stakeholders about this technology is the safety aspect regarding the transmission of Wolbachia to other species and the possibility that humans will contract Wolbachia. This study aimed to address these concerns, namely to find out whether horizontal transmission of Wolbachia occurred from A. aegypti that were released to other species and whether residents living in the released areas were infected with Wolbachia. The research was conducted in Dusun Nogotirto and Dusun Kronggahan (Sleman Regency), as well as in Dusun Jomblangan and Dusun Singosaren (Bantul Regency), Yogyakarta Special Province. Wolbachia qPCR screening using the target gene WD0513 was performed on 922 Culex quinquefasciatus Say and 331 Aedes albopictus (Skuse). ELISA test was carried out on 190 pairs of plasma samples, namely the sample before the Wolbachia frequency was established (still 80%). The results showed no evidence of Wolbachia transfer from Wolbachia-infected A. aegypti to other mosquito species coexisting in the same habitat or to humans. This study corroborates the safety evidence of Wolbachia-infected A. aegypti technology as an alternative to control dengue virus transmissio

Systematic analysis of factors that improve the efficiency and fidelity of CRISPR-directed gene integration

Abstract not currently available

Designing hybrid CRISPR-Cas12 and LAMP detection systems for treatment-resistant Plasmodium falciparum with in silico method

Genes associated with drug resistance of first line drugs for Plasmodium falciparum have been identified and characterized of which three genes most commonly associated with drug resistance are P. falciparum chloroquine resistance transporter gene (PfCRT), P. falciparum multidrug drug resistance gene 1 (PfMDR1), and P. falciparum Kelch protein K13 gene (PfKelch13). Polymorphism in these genes could be used as molecular markers for identifying drug resistant strains. Nucleic acid amplification test (NAAT) along with DNA sequencing is a powerful diagnostic tool that could identify these polymorphisms. However, current NAAT and DNA sequencing technologies require specific instruments which might limit its application in rural areas. More recently, a combination of isothermal amplification and CRISPR detection system showed promising results in detecting mutations at a nucleic acid level. Moreover, the Loop-mediated isothermal amplification (LAMP)-CRISPR systems offer robust and straightforward detection, enabling it to be deployed in rural and remote areas. The aim of this study was to develop a novel diagnostic method, based on LAMP of targeted genes, that would enable the identification of drug-resistant P. falciparum strains. The methods were centered on sequence analysis of P. falciparum genome, LAMP primers design, and CRISPR target prediction. Our designed primers are satisfactory for identifying polymorphism associated with drug resistant in PfCRT, PfMDR1, and PfKelch13. Overall, the developed system is promising to be used as a detection method for P. falciparum treatment-resistant strains. However, optimization and further validation the developed CRISPR-LAMP assay are needed to ensure its accuracy, reliability, and feasibilit

Prevalence and Distribution of Dengue Virus in <i>Aedes aegypti </i>in Yogyakarta City before Deployment of Wolbachia Infected <i>Aedes aegypti</i>

Indonesia is one of the countries where dengue infection is prevalent. In this study we measure the prevalence and distribution of dengue virus (DENV) DENV-infected Aedes aegypti in Yogyakarta City, Indonesia, during the wet season when high dengue transmission period occurred, as baseline data before implementation of a Wolbachia-infected Aedes aegypti trial for dengue control. We applied One-Step Multiplex Real Time PCR (RT-PCR) for the type-specific-detection of dengue viruses in field-caught adult Aedes aegypti mosquitoes. In a prospective field study conducted from December 2015 to May 2016, adult female Aedes aegypti were caught from selected areas in Yogyakarta City, and then screened by using RT-PCR. During the survey period, 36 (0.12%) mosquitoes from amongst 29,252 female mosquitoes were positive for a DENV type. In total, 22.20% of dengue-positive mosquitoes were DENV-1, 25% were DENV-2, 17% were DENV-3, but none were positive for DENV-4. This study has provided dengue virus infection prevalence in field-caught Aedes aegypti and its circulating serotype in Yogyakarta City before deployment of Wolbachia-infected Aedes aegypti