Electrochemical DNA Biosensor Based on Mercaptopropionic Acid-Capped ZnS Quantum Dots for Determination of the Gender of Arowana Fish.

A new electrochemical DNA biosensor based on mercaptopropionic acid (MPA)-capped ZnS quantum dots (MPA-ZnS QDs) immobilization matrix for covalent binding with 20-base aminated oligonucleotide has been successfully developed. Prior to the modification, screen-printed carbon paste electrode (SPE) was self-assembled with multilayer gold nanoparticles (AuNPs) and cysteamine (Cys). The inclusion of MPA-ZnS QDs semiconducting material in modified electrodes has enhanced the electron transfer between the SPE transducer and DNA leading to improved bioanalytical assay of target biomolecules. Electrochemical studies performed by cyclic voltammetry (CV) and differential pulsed voltammetry (DPV) demonstrated that the MPA-ZnS QDs modified AuNPs electrode was able to produce a lower charge transfer resistance response and hence higher electrical current response. Under optimal conditions, the immobilized synthetic DNA probe exhibited high selectivity towards synthetic target DNA. Based on the DPV response of the reduction of anthraquinone monosulphonic acid (AQMS) redox probe, the MPA-ZnS QDs-based electrochemical DNA biosensor responded to target DNA concentration from 1 × 10-9 µM to 1 × 10-3 µM with a sensitivity 1.2884 ± 0.12 µA, linear correlation coefficient (R2) of 0.9848 and limit of detection (LOD) of 1 × 10-11 µM target DNA. The DNA biosensor exhibited satisfactory reproducibility with an average relative standard deviation (RSD) of 7.4%. The proposed electrochemical transducer substrate has been employed to immobilize the aminated Arowana fish (Scleropages formosus) DNA probe. The DNA biosensor showed linearity to target DNA from 1 × 10-11 to 1 × 10-6 µM (R2 = 0.9785) with sensitivity 1.1251 ± 0.243 µA and LOD of 1 × 10-11 µM. The biosensor has been successfully used to determine the gender of Arowana fish without incorporating toxic raw materials previously employed in the hazardous processing conditions of polypyrrole chemical conducting polymer, whereby the cleaning step becomes difficult with thicker films due to high levels of toxic residues from the decrease in polymerization efficacy as films grew.

A simple optical pH sensor based on pectin and Ruellia tuberosa L-derived anthocyanin for fish freshness monitoring.

A simple optical pH sensor using the active compound anthocyanin (ACN), derived Ruellia tuberosa L. flower immobilized in a pectin membrane matrix, was been fabricated and employed to monitor the freshness of tilapia fish at room temperature and 4 oC storage. The optimum pectin weight and ACN concentrations were 0.1% and 0.025 mg/L. The sensor showed good sensitivity at 0.03 M phosphate buffer solution. The sensor's reproducibility was evaluated using 10 replicate sensors where a standard deviation of 0.045 or relative standard deviation of 9.15 was achieved. The sensor displayed an excellent response after 10 minutes of exposure, possessing a response stability for 10 consecutive days. The decrease in pH value of the Tilapia fish from 7.3 to 5 was observed in a 48 hour test, which can be used as the parameter when monitoring fish freshness.

Optical pH Sensor Based on Immobilization Anthocyanin from Dioscorea alata L. onto Polyelectrolyte Complex Pectin-Chitosan Membrane for a Determination Method of Salivary pH.

A simple optical pH sensor based on immobilization, Dioscorea alata L. anthocyanin methanol extract, onto a pectin-chitosan polyelectrolyte complex (pectin-chitosan PEC), has been successfully fabricated. The optical pH sensor was manufactured as a membrane made of pectin-chitosan PEC and the extracted anthocyanin. This sensor has the highest sensitivity of anthocyanin content at 0.025 mg/L in phosphate buffer and 0.0375 mg/L in citrate buffer. It also has good reproducibility with a relative standard deviation (%RSD) of 7.7%, and gives a stable response at time values greater than 5 min from exposure in a buffer solution, and the sensor can be utilized within five days from its synthesis. This optical pH sensor has been employed to determine saliva pH of people of different ages and showed no significant difference when compared to a potentiometric method.

Deep sequencing and in silico analysis of small RNA library reveals novel miRNA from leaf Persicaria minor transcriptome.

In current era, majority of microRNA (miRNA) are being discovered through computational approaches which are more confined towards model plants. Here, for the first time, we have described the identification and characterization of novel miRNA in a non-model plant, Persicaria minor (P. minor) using computational approach. Unannotated sequences from deep sequencing were analyzed based on previous well-established parameters. Around 24 putative novel miRNAs were identified from 6,417,780 reads of the unannotated sequence which represented 11 unique putative miRNA sequences. PsRobot target prediction tool was deployed to identify the target transcripts of putative novel miRNAs. Most of the predicted target transcripts (mRNAs) were known to be involved in plant development and stress responses. Gene ontology showed that majority of the putative novel miRNA targets involved in cellular component (69.07%), followed by molecular function (30.08%) and biological process (0.85%). Out of 11 unique putative miRNAs, 7 miRNAs were validated through semi-quantitative PCR. These novel miRNAs discoveries in P. minor may develop and update the current public miRNA database.

Anatomical and histological changes of uterine horn of Aceh cattle with repeat breeding.

OBJECTIVE: This study aimed to observe the changes in anatomical pathology, histopathology, and histomorphometry of uterine horn in Aceh cattle with repeat breeding (RB). MATERIALS AND METHODS: In this study, five uterine horns were collected from fertile Aceh cattle (N1, N2, N3, N4, and N5) and Aceh cattle with RB (RB1, RB2, RB3, RB4, and RB5). Changes in the anatomical pathology of uterine horn were observed prior to histopathological preparations. The results were analyzed descriptively. RESULTS: The observation of anatomical pathology revealed that the uterus of fertile cattle was yellowish white with horn-like shape consisting of hollow muscles, while RB cattle were reddish-pale. Furthermore, uterine horn swelling with purulent exudate was observed in the uterine horn of RB1 and RB4, while swelling with serous exudates was found in RB2, RB3, and RB5. Thin-walled uterine horn with caruncular atrophy was observed only in RB3. Histopathological observations showed erosion and hyperplasia of endometrial columnar epithelial and uterine glands in RB1, RB2, RB4, and RB5. Atrophy of uterine gland was found in RB3 and edema of caruncular was observed in RB1 and RB4. In addition, hemorrhage and inflammatory cell infiltration (neutrophils, lymphocytes, and macrophages) were found in all RB cattle. Histologically, cow uterus is divided into three layers, endometrium, myometrium, and perimetrium with the uterine gland found in the endometrium. Histomorphometric measurements found that the uterine horn wall (endometrial, myometrial, and perimetrium) of RB cattle were thicker than of fertile Aceh cattle, 208.06 ± 39.90 vs. 187.39 ± 29.09 µm, 400.138 ± 51.96 vs. 277.91 ± 42.88 µm, and 23.59 ± 9.67 vs. 18.53 ± 4.40 µm. However, the endometrial gland diameter of RB cattle is smaller than that of fertile Aceh cattle, 4.04 ± 0.88 vs. 4.99 ± 1.37 µm. CONCLUSION: The changes in anatomical pathology and histopathology of the uterine horn of Aceh cattle with RB indicate endometritis which caused by subacute and chronic bacterial infections. Endometritis causes disrupted blood circulation, which is characterized by hemorrhage, edema, and thickening of the endometrium, myometrium, and perimetrium, resulting in an increased diameter of the endometrial gland in RB Aceh cattle.

Small RNA-seq analysis in response to methyl jasmonate and abscisic acid treatment in Persicaria minor.

Persicaria minor (Kesum) is an important medicinal plant with high level of secondary metabolite contents, especially, terpenoids and flavonoids. Previous studies have revealed that application of exogenous phytohormone could increase secondary metabolite contents of the plant. MicroRNAs (miRNAs) are small RNAs that play important regulatory roles in various biological processes. In order to explore the possible role of miRNA in the regulation of these phytohormones signaling pathway and uncovering their potential correlation, we, for the first time, have generated the smallRNA library of Kesum plant. The library was developed in response to methyl jasmonate (MJ) and abscisic acid (ABA) treatment by using next-generation sequencing technology. Raw reads have been deposited to SRA database with the accession numbers, SRX2655642 and SRX2655643 (MJ-treated), SRXSRX2655644 and SRX2655645 (ABA-treated) and SRX2655646and SRX2655647 (Control).

Small RNA sequencing for secondary metabolite analysis in Persicaria minor.

Persicaria minor (kesum) is an important medicinal plant and commonly found in southeast countries; Malaysia, Thailand, Indonesia, and Vietnam. This plant is enriched with a variety of secondary metabolites (SMs), and among these SMs, terpenoids are in high abundance. Terpenoids are comprised of many valuable biomolecules which have well-established role in agriculture and pharmaceutical industry. In P. minor, for the first time, we have generated small RNAs data sets, which can be used as tool in deciphering their roles in terpenoid biosynthesis pathways. Fungal pathogen, Fusarium oxysporum was used as elicitor to trigger SMs biosynthesis in P. minor. Raw reads and small RNA analysis data have already been deposited at GenBank under the accessions; SRX2645684 (Fusarium-treated), SRX2645685 (Fusarium-treated), SRX2645686 (mock-infected), and SRX2645687 (mock-infected).

MicroRNA and Transcription Factor: Key Players in Plant Regulatory Network.

Recent achievements in plant microRNA (miRNA), a large class of small and non-coding RNAs, are very exciting. A wide array of techniques involving forward genetic, molecular cloning, bioinformatic analysis, and the latest technology, deep sequencing have greatly advanced miRNA discovery. A tiny miRNA sequence has the ability to target single/multiple mRNA targets. Most of the miRNA targets are transcription factors (TFs) which have paramount importance in regulating the plant growth and development. Various families of TFs, which have regulated a range of regulatory networks, may assist plants to grow under normal and stress environmental conditions. This present review focuses on the regulatory relationships between miRNAs and different families of TFs like; NF-Y, MYB, AP2, TCP, WRKY, NAC, GRF, and SPL. For instance NF-Y play important role during drought tolerance and flower development, MYB are involved in signal transduction and biosynthesis of secondary metabolites, AP2 regulate the floral development and nodule formation, TCP direct leaf development and growth hormones signaling. WRKY have known roles in multiple stress tolerances, NAC regulate lateral root formation, GRF are involved in root growth, flower, and seed development, and SPL regulate plant transition from juvenile to adult. We also studied the relation between miRNAs and TFs by consolidating the research findings from different plant species which will help plant scientists in understanding the mechanism of action and interaction between these regulators in the plant growth and development under normal and stress environmental conditions.