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The case of foodborne diseases has now become a serious global problem. Foodborne diseases occur due to the contamination of pathogenic bacteria. One of the pathogenic bacteria that cause foodborne diseases is Yersinia enterocolitica. This research aimed to determine the annealing temperature of Yersinia enterocolitica ymoA gene primers using the PCR method to ensure that the primers can amplify ymoA gene fragments in Yersinia enterocolitica. In this research, the concentration of DNA as a template used was 70.5 ng/µL with a purity of A260/280 1.88. Optimization of the annealing temperature of Yersinia enterocolitica ymoA gene primers using the Gradient PCR in a range of temperatures between 53°C - 62°C based on the melting temperature value of the ymoA primers. The results of the primers design of the ymoA gene were confirmed to amplify the DNA fragment of Yersinia enterocolitica with an amplicon length of 185 bp. Based on the results, the temperature of 60°C was chosen as the optimum primer annealing temperature for the ymoA gene in Yersinia enterocolitica because only a brightness band was produced from the electrophoresis. This result can be continued with primers testing the ymoA gene on meat samples using the Real-Time PCR method. [ABSTRACT FROM AUTHOR]

Vibrio parahaemolyticus must be detected quickly and precisely. This study uses the Gradient Polymerase Chain Reaction (PCR) method to evaluate the primer that can produce an amplicon at the ideal temperature for the Vibrio parahaemolyticus ToxR gene. The ToxR primer pair could amplify Vibrio parahaemolyticus ToxR fragments to form an amplicon with size 171 base pairs, according to the in-silico analysis conducted for the primer design. Gradient PCR was used to assess the primer's suitability for the design outcome and determine the ideal temperature to generate amplicons of the appropriate size with temperatures ranging from 53 to 62°C. The purity (A260/280) of the Vibrio parahaemolyticus isolate used as a template is 1.92, and its concentration is 51 ng/µl. The study's findings kindly reveal that annealing temperatures of 58°C produce the best outcomes for the ToxR primer, which results in the formation of a clear DNA band with a thickness band intensity and a size of 171 base pairs. Additionally, the primers and ideal temperatures discovered were utilized for the Real-Time PCR stage in creating sensitive, targeted, and quicker tools for detecting Vibrio parahaemolyticus in Seafood. [ABSTRACT FROM AUTHOR]

Listeria monocytogenes is a bacteria that causes food poisoning and is found in various types of food. It can infect humans with a mortality rate of up to 30%. This study aims to determine the potential of the internalin A (inlA) gene with Gradient Polymerase Chain Reaction (PCR) as a basis for developing a detection method for Listeria monocytogenes in vegetables using Real-Time PCR. This study obtained a round white colony on the Brain Heart Infusion (BHI) agar. In liquid media, the OD600 value is 1.418, indicating the bacteria's growth. The DNA used as a template had a concentration of 72 ng/µL with A260/280 purity of around 1.802. Variation of annealing temperature in Gradient PCR between 53 – 62°C based on melting temperature of the primer. The primer successfully amplifies the inlA gene fragment with an amplicon size 161 base pair. In addition, the optimum annealing temperature for these primers was 55-60°C based on the bright band and a single band formed in electrophoresis. In the future, primers and optimum annealing temperature can be used to develop a detection kit for Listeria monocytogenes bacteria in vegetables using the Real-Time PCR method. [ABSTRACT FROM AUTHOR]

Cronobacter sakazakii is a pathogen that causes necrotizing enterocolitis, sepsis, and meningitis in infants or neonates, with reported case-fatality rates varying from 40 to 80 percent. Therefore, it is necessary to develop fast and accurate detection of C. sakazakii. This study aims to get the primer pairs and optimal annealing temperature for the Polymerase Chain Reaction (PCR) method to detect C. sakazakii. The glutaredoxin B (grxB) gene was found suitable for detecting Cronobacter sakazakii, and a primer pair based on the grxB gene sequence was designed and synthesized. The genomic DNA of Cronobacter sakazakii strain ATCC 29544 was isolated and then used as a template for PCR assay. Optimization of running PCR conditions was carried out by varying the annealing temperature at 53-62°C, and the template DNA concentration of C. sakazakii is 53 ng/µL with a purity (A260/280) of 1,853. The result showed that the primer grxB gave optimal results at the annealing temperature range between 57-61°C and could amplify the grxB C. sakazakii fragment to produce an amplicon with a size of 151 base pairs (bp), and only a single band was formed. Furthermore, the primer design results and the optimum temperature obtained will be used to develop a sensitive and specific rapid detection method of C. sakazakii in food samples using real-time PCR. [ABSTRACT FROM AUTHOR]

Coumarin is a therapeutic agent, found naturally as a secondary metabolite in plants, bacteria, fungi, and essentialoils, and can be chemically synthesized. The Knoevenagel reaction is one of the most useful C=C bond formation methodsinvolving the condensation between benzaldehyde and activated methylenes in the presence of an amine. This study aims to synthesize coumarin with microwave-assisted via Knoevenagel condensation. This study synthesized coumarin from 2-hydroxybenzaldehyde and various activated methylenes with various catalysts. The mixture was irradiated in the microwave for several reaction times (60 s, 70 s, 80 s, 10 minutes, and 15 minutes) at 5-10 intervals. TLC monitored the reaction using n-hexane and ethyl acetate (8:2). The product was purified by recrystallization. The product was tested for purity by TLC and melting point, followed by structural elucidation by UV-Vis spectrophotometry. It was found that the optimal time for 3-acetylcoumarin synthesis was 60 s and 10 minutes for ethyl coumarin 3-carboxylate. [ABSTRACT FROM AUTHOR]

Chalcone compound is one of the derivatives of flavonoids and is reported to have many bioactivities such as antibacterial, anticancer, antioxidant, and anti-inflammatory. The use of chalcone continuously supports research on its synthesis and bioactivity. Microwave-assisted organic synthesis is used because it reduces reaction times, makes easy work, and produces cleaner products. A series of chalcones from vanillin with acetophenone and 4-hydoxyacetophenone were synthesized by the Claisen-Schmidt reaction using a microwave using a KOH base catalyst. The mixtures were irradiated by microwave for 5 minutes. After the reaction, cold distilled water and HCl were added to the mixtures and then left for 24 hours in the refrigerator, after which it was recrystallized. Pure chalcones were obtained with yields of 97.64 % and 80.65 %. The melting point of 80°C-84°C and 86°C-90°C. UV Spectrophotometry was carried out to characterize the obtained chalcones. [ABSTRACT FROM AUTHOR]

The development of rapid, sensitive, and specific detection methods has increasingly become the attention in handling food poisoning cases. In this study, the method of Real-Time PCR (RT-PCR) with nuc gene target was developed for the detection of Staphylococcus aureus in food samples of meat and milk. The results showed that the primer nuc gene with Gradient PCR could detect Staphylococcus aureus bacteria from the pure culture at 57-61 oC by producing amplicons measuring 135 base pairs. Amplification by RT-PCR in the same sample at an annealing temperature of 58oC and a concentration of 50 ng gave a Ct value of 18.88-19.56. The primer specificity test of that gene shows that the primer can recognize the target bacteria with a strong fluorescent signal at the Melting Curve of 79.02 oC, whereas non-target bacteria give low signals and different Tm values. The primer sensitivity of the nuc gene has a detection limit (LOD) of 98.4 pg/µL or 8.8 x 102 CFU/mL. Based on these results, the RT PCR method with the nuc gene has the potential to be a method of detecting specific, sensitive, and rapid for Staphylococcus aureus bacteria in a food sample. [ABSTRACT FROM AUTHOR]

The development of technology and information is already well-known in the Industrial Revolution 4.0. It brings a very large impact on education in this century. The use of technology can be effectively better in learning media. The study focused to develop an electronic module (E-Module) by using the Flip PDF Professional application as a medium for learning organic chemistry that concern about carbohydrates that can be accessed through smartphones on the Android platform, desktops, and laptops. The E-Module carbohydrates were developed with the Chemistry undergraduate program curriculum. The contents of the E-module consist of carbohydrate discussion material which is equipped with video, pictures, hyperlinks, flash, and formative tests on each sub-chapter material. The research method used was Research and Development (R&D) with the Borg and Gall model. The E-Module development procedure was carried out by analyzing initial needs in the form of interviews with lecturers and distributing questionnaires to 75 chemistry students. Furthermore, the E-module that has been compiled is validated by material, language, and media experts using a standard Likert scale instrument. Based on the assessment of material, language, and media, it can't be ignored that this E-module gets a good interpretation so that it is suitable for use in the learning process in the organic chemistry course with carbohydrate ingredients. The E-Module dissemination in the organic chemistry learning process is being carried out and will be published in the next article. [ABSTRACT FROM AUTHOR]

Detection of food-borne bacterial pathogens was developed to overcome the limitations. The aim of this research was to develop Salmonella typhimurium detection by Real Time Polymerase Chain Reaction (RT-PCR) using two pairs of primers. The ability of primer pairs to detect S. typhimurium is seen from cycle threshold or Ct. Artificially contaminated milk sample with 24 ng each microliter can be detected with fljB (flagellin gene) primers on Ct 12,933 and with STM4497 (hypothetical protein code) primers on Ct 13,665. The specificity test of both primers showed that melting temperature (Tm) of fljB was 80,5 degree Celsius, and STM449 was 81,6 degree Celsius. FljB and STM4497 primers gave an average detection limit respectively of 11,75 Colony Forming Unit (CFU) each milliliter and 6,8 CFU each milliliter. The time needed throughout the detection process of S. typhimurium with fljB and STM4497 primers is faster than conventional methods. Based on the results it can be concluded that primers fljB and STM449 S. typhimurium can be applied to detection and quantification of S. typhimurium in milk samples. [ABSTRACT FROM AUTHOR]

Malignant tumors are one of the main causes of death in the world. Until now the search for cytotoxic (antitumor) compounds from nature, particularly from plants, is being a continuation activities. One group of plants that produce potential cytotoxic compounds is the Cryptocarya, one of the large genera of the Lauraceae family. As a part of our chemical and cytotoxic evaluation of the Cryptocarya species, we examined three species of Indonesian Cryptocarya. The sample of the wood of C. konishii hayata was collected from Cibodas Botanical Garden, West Java while the stem bark of C. phoebeopsis and C. cagayanensis were obtained from Sorong, Papua. Our investigation of flavonoid constituents on these species afforded three chalcone compounds i.e. desmethylinfectocaryone (1), infectocaryone (2) and cryptocaryone (3). The molecular structures of the isolated compounds were determined based on spectroscopic data, including UV, IR, 1D and 2D NMR. Cytotoxic effects of the compounds were evaluated using MTT [3-(4,5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide] assay. Compound 1, 2 and 3 displayed strong cytotoxic properties (IC50 < 2 µg/mL) against Murine Leukemia P388 and HL 60 (blood premyelocytic leukemia) cells whereas 2 and 3 exhibited strong cytotoxicity properties against HCT116 (colon cancer). Cryptocaryone (3) also showed moderate cytotoxic properties (IC50 < 10 µg/mL) towards A549 (human lung adenocarcinoma epithelial) cells. [ABSTRACT FROM AUTHOR]

There is a lot of public concern over food safety. Food-safety cases recently, including many food poisoning cases in both the developed and developing countries, considered to be the national security threats which involved police investigation. Quick and accurate detection methods are needed to handle the food poisoning cases with a big number of sufferers at the same time. Therefore, the research is aimed to develop a specific, sensitive, and rapid result molecular detection tool for foodborne pathogen bacteria. We, thus, propose genomic level approach with Polymerase Chain Reaction. The research has successfully produced a specific primer to perform amplification to fim-C S. typhi, E. coli, and pef Salmonella typhimurium genes. The electrophoresis result shows that amplification products are 95 base pairs, 121 base pairs, and 139 base pairs; and all three genes are in accordance with the size of the in silico to third genes bacteria. In conclusion, the research has been successfully designed a specific detection tool to three foodborne pathogen bacteria genes. Further stages test and the uses of Real-time PCR in the detection are still in the trial process for better detection method. [ABSTRACT FROM AUTHOR]