Improving siRNA design targeting nucleoprotein gene as antiviral against the Indonesian H5N1 virus

BackgroundSmall interfering RNA technology has been considered a prospective alternative antiviral treatment using gene silencing against influenza viruses with high mutations rates. On the other hand, there are no reports on its effectiveness against the highly pathogenic avian influenza H5N1 virus isolated from Indonesia.ObjectivesThe main objective of this study was to improve the siRNA design based on the nucleoprotein gene (siRNA-NP) for the Indonesian H5N1 virus.MethodsThe effectiveness of these siRNA-NPs (NP672, NP1433, and NP1469) was analyzed in vitro in Marbin-Darby canine kidney cells.ResultsThe siRNA-NP672 caused the largest decrease in viral production and gene expression at 24, 48, and 72 h post-infection compared to the other siRNA-NPs. Moreover, three serial passages of the H5N1 virus in the presence of siRNA-NP672 did not induce any mutations within the nucleoprotein gene.ConclusionsThese findings suggest that siRNA-NP672 can provide better protection against the Indonesian strain of the H5N1 virus.     

Update on Non-Alcoholic Fatty Liver Disease-Associated Single Nucleotide Polymorphisms and Their Involvement in Liver Steatosis,Inflammation, and Fibrosis: A Narrative Review

Genetic factors are involved in the development, progression, and severity of NAFLD. Polymorphisms in genes regulating liver functions may increase liver susceptibility to NAFLD. Therefore, we conducted this literature study to present recent findings on NAFLD-associated polymorphisms from published articles in PubMed from 2016 to 2021. From 69 selected research articles, 20 genes and 34 SNPs were reported to be associated with NAFLD. These mutated genes affect NAFLD by promoting liver steatosis (PNPLA3, MBOAT7, TM2SF6, PTPRD, FNDC5, IL-1B, PPARGC1A, UCP2, TCF7L2, SAMM50, IL-6, AGTR1, and NNMT), inflammation (PNPLA3, TNF-α, AGTR1, IL-17A, IL-1B, PTPRD, and GATAD2A), and fibrosis (IL-1B, PNPLA3, MBOAT7, TCF7L2, GATAD2A, IL-6, NNMT, UCP, AGTR1, and TM2SF6). The identification of these genetic factors helps to better understand the pathogenesis pathways of NAFLDKey Words: Fibrosis, Inflammation, Non-alcoholic fatty liver disease, Polymorphism     

Influence of weed management measures on glyphosate resistance and endophyte infection in naturalized Italian ryegrass (Lolium multiflorum)

Recently, glyphosate‐resistant Italian ryegrass (Lolium multiflorum Lam.) was found on rice paddy levees in a western region of Shizuoka Prefecture, Japan. Naturalized populations of Italian ryegrass are frequently infected with fungal Epichloë endophytes. Endophytes often confer benefits to their host grasses. This study investigated the influence of five weed management treatments on glyphosate resistance and endophyte infection in Italian ryegrass that was growing on paddy levees where glyphosate‐resistant individuals were dominant. The weed management treatments were: (i) mowing once before the grass flowered; (ii) mowing once during flowering; (iii) mowing twice during flowering; (iv) glyphosate application before flowering; and (v) no treatment. The seeds were collected from the treatment plots in 2013 and 2014. The seeds were examined for endophyte infection and the seedlings that had been grown from the seeds were tested for the frequency of glyphosate resistance. The seedlings that had been derived from the glyphosate treatment showed higher frequencies of glyphosate resistance than those seedlings that had been derived from all the other treatments. Endophytes were found in all populations of the seeds from the paddy levees, with higher infection rates in the seeds that had been derived from the glyphosate treatment and the twice‐mowed treatment. There was a significant relationship between the endophyte infection frequency in the seeds and glyphosate resistance in the seedlings that had been grown from the same populations. The results indicate that where glyphosate herbicides are frequently used, selection for glyphosate‐resistant Italian ryegrass occurs, and along with this, the frequency of endophyte infection also increases.     

Comparative characteristic study from bone marrow-derived mesenchymal stem cells

Tissue engineering has been extensively investigated and proffered to be a potential platform for novel tissue regeneration. The utilization of mesenchymal stem cells (MSCs) from various sources has been widely explored and compared. In this regard, MSCs derived from bone marrow have been proposed and described as a promising cell resource due to their high yield of isolated cells with colony-forming potential, self-renewal capacity, MSC surface marker expression, and multi-lineage differentiation capacities in vitro. However, there is evidence for bone marrow MSCs (BM-MSCs) both in vitro and in vivo from different species presenting identical and distinct potential stemness characteristics. In this review, the fundamental knowledge of the growth kinetics and stemness properties of BM-MSCs in different animal species and humans are compared and summarized. Finally, to provide a full perspective, this review will procure results of current information studies focusing on the use of BM-MSCs in clinical practice.     

Genotypic characterization of Staphylococcus aureus isolated from bovines, humans, and food in Indonesia

The present study determined the genetic relationships between 41 Staphyloccocus (S.) aureus isolates from bovines, humans, and food using a single enzyme amplified fragment length polymorphism (AFLP) technique. We evaluated the prevalence of staphylococcal enterotoxin (SE) genes and other virulence gene determinants by PCR. The identification of S. aureus was based on culturing and biochemical tests, and by amplifying a specific section of the 23S rRNA gene. PCR amplification of the SE genes (sea, seb, sec, see, seg, seh, and sei) singly or in combination was observed. Most isolates of bovine origin harbored hla (84%) and cap5 (74%), while most isolates from humans harbored hla (73%), cap8 (91%), and fnbA (100%). Strains from food sources were positive for hla (100%), cap5 (100%), and cap8 (64%) unlike isolates from humans or bovines. A single enzyme AFLP analysis revealed a correlation between AFLP clusters of some strains and the source of the isolates The genotypic results of the present study might help to better understand the distribution of prevalent S. aureus clones among humans, bovines, and food and will help control S. aureus infections in Indonesia.     

Preparation of Dried Culture of Probiotic Rhizopus oryzae in Two Different Mediums as Feed Additive to Indigenous Chicken

Fungus Rhizopus oryzae has a probiotic potential, but the potency is not yet explored. Probiotics are usually used as feed additive, including to the chicken. Probiotic needs a suitable medium for their life. The aim of the study was to know which preparations can produce suitable medium for the growth of Rhizopus oryzae on the dried culture formed. The study used two preparations. Preparation 1 （a） consisted of rice bran （1 kg） and culture ofRhizopus oryzae on Potato Dextrose Broth （PDB medium） （500 mL）; then preparation 1 （b） consisted of corn grain （1 kg） and culture of Rhizopus oryzae on PDB medium （500 mL）. Preparation 2 （a） consisted of rice bran （1 kg）, culture of Rhizopus oryzae on Potato Dextrose Agar （PDA medium） （seven petri dishes） and was added with sterile distilled water （500 mL）; preparation 2 （b） consisted of corn grain （1 kg）, culture of Rhizopus oryzae on PDA medium （seven petri dishes） and was added with sterile distilled water （500 mL）. The culture of Rhizopus oryzae even both on PDB and PDA were 3 d of incubation. All preparations were incubated for 7 d then they were dried under the sun. Parameter evaluation was the growth performance of the fungus in the dried culture media. The growth performance was analyzed by microscopic analysis and the number of colony of the fungi in the two dried culture media. The results showed that from the macroscopic analysis, the sample of both dried culture media which were placed on PDA medium showed the growth of mycelia and spores of the same kind of fungus and it was proved by microscopic observation that the fruiting bodies （mycelia and spores） of that ftmgus was Rhizopus oryzae. The colonies number of the fungi from preparation 1 （a） and 1 （b） were 0,4 × 10 cfu/gr and 2.4 × 10^3 cfu/gr, respectively, and from preparation 2 （a） and 2 （b） were 1.3 × 10^5 cfu/gr and 8.3 ×10^2 cfu/gr, respectively. It could be concluded that the product of the dried culture of probiotic     

Morphological characterization of Horsfield’s treeshrew Tupaia javanica lingual papillae: Light microscopy and scanning electron microscopy studies

The Horsfield's treeshrew (Tupaia javanica) is one of the shrews found on the island of Java that mainly feeds on fruits and occasionally insects. This shrew has an important role in the ecosystem as it spreads plant seeds and controls insects. To this day, there is no available knowledge about the structure of Horsfield's treeshrew's tongue. Therefore, the aim of this study was to provide data on the tongue structure of the Horsfield's treeshrew through two different methods. We investigated the tongue morphology and lingual papillae distribution of Tupaia javanica using 1) scanning electron microscopy (SEM) for microscopic structure and 2) histology for microscopic organization. Our macroscopic results revealed that the tongue of Tupaia javanica can be divided into three parts: apex, corpus and radix. The dorsal surface of the medial corpus is the median groove extending to the radix. Observation using SEM and light microscopy (LM) showed that the apex of the tongue bears three types of papillae: scale-like filiform, small filiform and fungiform. These are rosette filiform papillae, cornflower filiform papillae, conical papillae and fungiform papillae in the corpus. Meanwhile, the radix contains small filiform papillae, scale-like filiform papillae, vallate papillae, conical papillae and fungiform papillae. On the lateral posterior radix, Weber's gland reacts positively to PAS reaction. Taking together, our research shows that there are variations in the papillae on the tongue surface of Horsfield's treeshrew, and the variations of these papillae are very dependent on the pattern, type of food and the function of the tongue.