Putative DNA-dependent RNA polymerase in Mitochondrial Plasmid of Paramecium caudatum Stock GT704

Mitochondria of Paramecium caudatum stock GT704 has a set of four kinds of linear plasmids with sizes of 8.2, 4.1, 2.8 and 1.4 kb. The plasmids of 8.2 and 2.8 kb exist as dimers consisting of 4.1- and 1.4-kb monomers, respectively. The plasmid 2.8 kb, designated as pGT704-2.8, contains an open reading frame encodes for putative DNA-dependent RNA polymerase (RNAP). This study reveals that this RNAP belongs to superfamily of DNA/RNA polymerase and family of T7/T3 single chain RNA polymerase and those of mitochondrial plasmid of fungi belonging to Basidiomycota and Ascomycota. It is suggested that RNAP of pGT704-2.8 can perform transcription without transcription factor as promoter recognition. Given that only two motifs were found, it could not be ascertained whether this RNAP has a full function independently or integrated with mtDNA in carrying out its function

DNA Barcoding of Sangihe Nutmeg (Myristica fragrans) using matK Gene

Nutmeg (family: Myristicaceae) is a plant that originated from Banda islands and is widely cultivated in several places in the world. Secondary metabolites of this plant have a high value because of their benefits for the health, food, and beauty industries. This study aims at developing DNA barcode for nutmeg (Myristica fragrans) using standard recommended fragment of matK (maturase K) gene. Universal matK primer pairs were used to amplify 889 bp DNA fragment. BLAST search from NCBI site showed that Sangihe nutmeg has 100% identity with Myristica fatua, M. maingayi, and M. globosa. It also has 3 nucleotides difference with Rivola sebifera (identity 99.58%) and 4 nucleotides difference with Knema laurina (identity 99.43%). It can be inferred from this study that single locus of matK gene cannot be used to differentiate species in Myristica; it can only be used to differentiate the genus level within family Myristicaceae

IDENTIFIKASI KOMPONEN SENYAWA DALAM EKSTRAK N-HEKSANA UMBI RUMPUT TEKI (Cyperus rotundus L.) DENGAN ANALISIS GC-MS

ABSTRACTNut grass (Cyperus rotundus L.) is a perennial weed which the tuber is used traditionally in treating several ailments. This study aimed to identify the phytochemical compounds from nut grass tuber. Extraction was carried out by maceration using n-hexane as solvent and then were analyzed with Gas Chromatography-Mass Spectrometry (GC-MS). The result showed that there were 48 peaks with 144 probable compounds detected in n-hexane extract of nut grass tuber. 7-Isopropenyl-1,4a-dimethyl-4,4a,5,6,7,8-hexahydro-3H-naphthalen-2-one; 1(2H)-Naphthalenone, 3,4,4a,5,6,7-hexahydro-4a,5-dimethyl-3-(1-methylethenyl)-, [3S-(3a,4aa,5a)]-; and 1,7-Dimethyl-4-(propan-2-ylidene)tricyclo[4.4.0.02,7]decan-3-one had the highest concentrations namely 43,92% in the extract. This study concluded that the compounds were classified as sesquiterpenoids which had various biological activities. Keywords: N-hexane extract, GC-MS, chemical compounds, Nut Grass (Cyperus rotundus L.).  ABSTRAKRumput teki (Cyperus rotundus L.) adalah gulma perenial yang umbinya dimanfaatkan secara tradisional oleh masyarakat untuk mengobati beberapa jenis penyakit. Penelitian ini bertujuan untuk mengidentifikasi senyawa-senyawa fitokimia dari umbi rumput teki.  Ekstraksi dilakukan dengan cara maserasi menggunakan pelarut n-heksana dan kemudian dilanjutkan dengan analisis Gas Chromatography-Mass Spectrometry (GC-MS). Hasil analisis menunjukkan terdapat 48 peak dengan 144 kemungkinan komponen senyawa yang terdeteksi dalam ekstrak n-heksana umbi rumput teki. Senyawa 7-Isopropenyl-1,4a-dimethyl-4,4a,5,6,7,8-hexahydro-3H-naphthalen-2-one; 1(2H)-Naphthalenone, 3,4,4a,5,6,7-hexahydro-4a,5-dimethyl-3-(1-methylethenyl)-, [3S-(3a,4aa,5a)]-; dan 1,7-Dimethyl-4-(propan-2-ylidene)tricyclo[4.4.0.02,7]decan-3-one memiliki persentase kadar tertinggi yaitu 43,92% dalam ekstrak. Berdasarkan hasil penelitian disimpulkan bahwa senyawa-senyawa ini termasuk dalam golongan seskuiterpenoid yang memiliki aktivitas biologis yang beragam. Kata Kunci: Ekstrak n-heksana, GC-MS, komponen senyawa, Rumput Teki (Cyperus rotundus L.

Mechanistic insights of Cucumis melo L. seeds for gastrointestinal muscle spasms through calcium signaling pathway–related gene regulation networks in WGCNA and in vitro, in vivo studies

Background: In addition to the nutritional benefits of Cucumis melo L., herbalists in Pakistan and India employ seeds to treat various ailments. This study aimed to determine the regulatory role of C. melo seeds in calciummediated smooth muscle contraction. Methods: We identified and quantified the phytochemicals of C. melo with LC ESI–MS/MS and HPLC, then conducted in vitro and in vivo tests to confirm the involvement in smooth muscle relaxation. Then, diarrheapredominant irritable bowel syndrome gene datasets from NCBI GEO were acquired, DEGs and WGCNA followed by functional enrichment analysis. Next, molecular docking of key genes was performed. Results: The quantification of C. melo seeds revealed concentrations of rutin, kaempferol, and quercetin were 702.38 μg/g, 686.29 μg/g, and 658.41 μg/g, respectively. In vitro experiments revealed that C. melo seeds had a dose-dependent relaxant effect for potassium chloride (80 mM)–induced spastic contraction and exhibited calcium antagonistic response in calcium dose-response curves. In in vivo studies, Cm.EtOH exhibited antidiarrheal, antiperistaltic, and antisecretory effects. The functional enrichment of WGCNA and DEGs IBS-associated pathogenic genes, including those involved in calcium-mediated signaling, MAPK cascade, and inflammatory responses. MAPK1 and PIK3CG were identified as key genes with greater binding affinity with rutin, quercitrin, and kaempferol in molecular docking. Conclusions: The bronchodilator and antidiarrheal effects of C. melo were produced by altering the regulatory genes of calcium-mediated smooth contractionUniversidade de Vigo/CISU

The potent antimicrobial spectrum of patchouli: systematic review of its antifungal, antibacterial, and antiviral properties

Intention towards natural essential oils from medicinal plants has increased rapidly over the past decade as these oils have antimicrobial and antioxidant properties against various chronic diseases. One essential oil source with antimicrobial properties is the essential oil from Pogostemon cablin (Blanco) Benth. This review aims to provide information on using patchouli oil as an antimicrobial against bacterial, fungal, and viral pathogens in the last five years. There were 37 articles found in the PUBMED database by June 15, 2023. After searching, 6 of them were duplicates. A total of 2 papers were inaccessible, 4 were not research articles, and fivewere excluded because they were irrelevant to the scope of this study. This review shows that research related to patchouli as an antimicrobial in the last five years involves Pogostemon cablinleaf samples as silver nanoparticle bioreductors. Patchoulioil is used in membrane, nanocomposite film, and starch hydrogel manufacturing. Patchouli oil is a prestigious antimicrobial agent because it can fight numerous pathogenic microbes from bacteria, fungi, and viruses

Genome editing and cancer: How far has research moved forward on CRISPR/Cas9?

Financiado para publicación en acceso aberto: Universidade de Vigo/CISUGCancer accounted for almost ten million deaths worldwide in 2020. Metastasis, characterized by cancer cell invasion to other parts of the body, is the main cause of cancer morbidity and mortality. Therefore, understanding the molecular mechanisms of tumor formation and discovery of potential drug targets are of great importance. Gene editing techniques can be used to find novel drug targets and study molecular mechanisms. In this review, we describe how popular gene-editing methods such as CRISPR/Cas9, TALEN and ZFNs work, and, by comparing them, we demonstrate that CRISPR/Cas9 has superior efficiency and precision. We further provide an overview of the recent applications of CRISPR/Cas9 to cancer research, focusing on the most common cancers such as breast cancer, lung cancer, colorectal cancer, and prostate cancer. We describe how these applications will shape future research and treatment of cancer, and propose new ways to overcome current challengesKing Khalid University in Abha, Saudi Arabia | Ref. RGP: 52/2/144

OVERPRODUCTION OF MERCURIC REDUCTASE FROM MERCURY-RESISTANT BACTERIA KLEBSIELLA PNEUMONIAE ISOLATE A1.1.1

Mercury is a highly toxic compound in human. It can, however, be detoxified by mercuric reductase (MerA) protein derived from mercury resistant bacteria. This study aims to obtaine MerA protein by transforming merA gene into Escherichia coli BL21. Nucleotide sequence of merA gene of mercury resistant bacteria Klebsiella pneumoniae isolates A111, optimized by using gene program designers (www.dna20/com) then commercially synthesized and cloned in pET32b expression plasmid vector. Plasmid was transformed into Escherichia coli BL21 to produce MerA protein recombinant, induced with isopropyl-β-D-thiogalactopyranoside (IPTG). MerA proteins were analyzed by 10% sodium dodecyl sulphate poly acrylamide gel electrophoresis (SDS PAGE). The result showed that MerA protein with 60kDa was detected on SDS PAGE. The obtained MerA protein can be used in further research for the enzymatic detoxification of inorganic mercury

OVERPRODUCTION OF MERCURIC REDUCTASE FROM MERCURY-RESISTANT BACTERIA KLEBSIELLA PNEUMONIAE ISOLATE A1.1.1

Mercury is a highly toxic compound in human. It can, however, be detoxified by mercuric reductase (MerA) protein derived from mercury resistant bacteria. This study aims to obtaine MerA protein by transforming merA gene into  Escherichia coli BL21. Nucleotide sequence of merA  gene of mercury resistant bacteria Klebsiella pneumoniae isolates A111, optimized by using gene program designers (www.dna20/com) then commercially synthesized and cloned in pET32b expression plasmid vector. Plasmid was transformed into Escherichia coli BL21 to produce MerA protein recombinant, induced with isopropyl-β-D-thiogalactopyranoside (IPTG). MerA proteins were analyzed by 10% sodium dodecyl sulphate poly acrylamide gel electrophoresis (SDS PAGE). The result showed that MerA protein with 60kDa was detected on SDS PAGE. The obtained MerA protein can be used in further research for the enzymatic detoxification of inorganic mercury.Key words: mercuric reductase, merA gene, MerA protein, Escherichia coli BL2

Sequence Analysis of the Cytochrome C Oxidase Subunit I Gene of Pseudagrion pilidorsum (Odonata: Coenagrionidae)

Pseudagrion pilidorsum is 1 of over 140 species of Pseudagrion (in the family Coenagrionidae), the largest genus of damselfly. This species exhibits dimorphism due to the different body colorations of males and females, making them difficult to distinguish from other congeneric species. This study analyzed the cytochrome C oxidase subunit I (COI) gene sequence of P. pilidorsum found in Bogani Nani Wartabone National Park (North Sulawesi) and compared it with other sequences of P. pilidorsum from distinct geographical locations in Asia. The COI gene for the Sulawesi specimen was amplified using the universal primer pair LCO1490 and HCO2198. A sequence homology search was conducted through BLAST. Multiple sequence alignment was executed using CLUSTAL O (1.2.1). A phylogenetic tree was constructed using the Neighbor-Joining method, and genetic distance was calculated using the Kimura 2-parameter. The COI gene sequence of the Sulawesi specimen lies in the range of 83.99-89.10% with other P. pilidorsum deposited at GenBank, namely KF369526 (Sarawak specimen), AB708543, AB708544, and AB708545 (Japan specimens). The genetic distance falls in the range of 0.146-0.149 between the Sarawak specimen and the Japan specimen; 0.122-0.125 between the Sulawesi and Japan specimens; and 0.185 between the Sulawesi specimen and the Sarawak specimen. It can thus be inferred that the Sarawak and Japan specimens may not belong to the same species; the Sulawesi and Japan specimens may not belong to the same species; and the Sarawak specimen and Sulawesi specimens might be placed in different genera.&nbsp