Journal of Applied Biotechnology Reports
Volume:6 Issue: 4, Autumn 2019

Folate, also known as vitamin B9, is essential in cell metabolism and very important especially for pregnant women and lactating mothers. Natural folate is available in food but it is very unstable. Synthetic folate is generally used as an alternative to meet daily needs due to its stability, even though it has a negative effect causing a variety of metabolic disorders. Some lactic acid bacteria have been reported as being able to synthesize natural folate during the fermentation process. Lactic acid bacteria are the main microorganisms for lactic fermentation such as fermented milk, fruits, and vegetables. Milk is the most nutritious food and contains folate-binding protein, hence it is considered the ideal fermentation medium to increase folate stability during storage. Fermentation of milk with folate-producing lactic acid bacteria can be used as a technique to produce natural folate-rich fermented foods as an attempt to prevent folate deficiency without side effects to the consumers.

Polymerase chain reaction–single-strand conformation polymorphism (PCR-SSCP) and PCR–restriction fragment length polymorphism (PCR-RFLP) are two independent methods used in the post-amplification genotyping of DNA variations. Both techniques are used in a wide range of screening applications to characterize single nucleotide polymorphisms (SNPs). The PCR-SSCP enables the identification of a potentially causative unknown SNP that could not be identified by PCR-RFLP. However, because complicated steps are not required to perform PCR-RFLP, it is used in many applications. On the other hand, PCR-RFLP is easier to process in terms of time and handover experience, the detection of a particular unknown SNP by PCR-SSCP has further chances. The simplicity of PCR-RFLP does not mean that it is better than PCR-SSCP. The reason is the limited ability of PCR-RFLP to detect nucleotide variations, which often go undetected because each restriction enzyme (RE) scans only a few recognition sequences, and other sequences are ignored. Furthermore, the efficacy of PCR-SSCP is sometimes hindered by many optimizations and also lack of experience. As PCR-SSCP allows other sequences within an amplicon to be separated and characterized, the choice between PCR-RFLP and PCR-SSCP is largely dependent on the reason for each genotyping experiment. This review provides a useful guide for comparing PCR-RFLP and PCR-SSCP in terms of their concepts, efficiency, ease of use, interpretation, and sensitivity as well as several other parameters. The comparison is extended to the practical applications of both techniques in terms of their utilization in molecular diagnostics and related applications.

Avocado and soya unsaponifiables (ASU) have been reported to be useful for the treatment of cartilage diseases. The aim of this study was to detect whether avocado/soybean can have any effect on the chondrogenesis of human adipose-derived stem cells on polylactic-co-glycolic acid/fibrin hybrid scaffold or not. The poly-lactic-co-glycolic acid (PLGA)/fibrin scaffolds were seeded with cultured human adipose tissue-derived stem cells (hADSCs), which were then divided into three groups: control, TGF-β3, and ASU and the results were analyzed 14 days later. The viability of the cells in different groups were assessed by MTT. The expression of chondrogenic-related genes Sox9, type II collagen, Aggrecan, type X collagen, and type I collagen were quantified by real time polymerase chain reaction (PCR). Protein expression levels of collagen type II and X were evaluated by Western blotting. Enhanced cellular viability was observed in the ASU group compared to the transforming growth factor beta-3 (TGF-β3) group. Analysis of aggrecan (Agg), type II collagen (Coll2) and SOX9 revealed that ASU and TGF-β3 induce hADSCs on PLGA/fibrin scaffold to differentiate into chondrocytes in-vitro. Moreover, a significant decrease was observed in the expression of type X (Coll10) and I collagen (Coll1) genes in the ASU group compared to the TGF-β3 group. Protein levels of type II collagen (Coll2) significantly increased in TGF-β3 and ASU groups in comparison with those of the control group. However, protein levels of Type X collagen (Coll10) significantly declined in the ASU group when compared with the TGF-β3 group. The results of the present study indicated that hADSCs containing the ASU in PLGA/fibrin hybrid scaffold are an effective way to potentially enhance Cartilage-specific genes with less hypertrophy and Fibrosis in-vitro.

Phytochemicals are one wide class of nutraceuticals found in plants which act as antioxidants. In this research, the essential oil (EO) of Teucrium polium L., Lamiaceae, collected from Mascara province, situated in the Algerian northwestern, where their chemical composition varies according to geographical origin, season variation, and climatic conditions were studied. The extraction of EO was performed by hydrodistillation. Then, the chemical compounds were identified by gaz chromatography coupled to a mass spectrometer (GC–MS). In parallel, the antioxidant activity was evaluated using the DPPH test. The yield of the EO of T. polium L. varied during different seasons with the highest in winter season, at vegetative stage (S1), while the same EO (S1) was significantly more efficient as an antioxidant than the EO harvasted at the flowering stage (S2) with IC50 values 3.90±0.05, 16.14±0.15 mg/mL, respectively (P < 0.001). These extracts are predominantly constituted by limonene (29.87%-26.39%), spathulenol (17.24%-13.29%), camphor (0.0%-8.20%), pinocarvone (7.76%-5.60%), tau-cadinol (5.41%-3.67%), pinene oxide (0.0%-4.78%), α-terpineol (0.0%-4.6%), 1-adamantanemethylamine (0.0%-9.80%) and β- myrcene (0.0%-4.02%). The results show that both EOs can be considered as potential sources of natural antioxidants. However, the vegetative stage was the best stage for harvesting the EO of T. polium L. which can be used as an alternative source of synthetic compounds.

Today, one of the most important challenges of the therapeutic system is the resistance of bacteria against different antibiotics especially in intensive care units which lead to an increase in hospitalization time and the patients’ expenses. Acinetobacter baumannii is one of the most significant contaminating bacteria in intensive care units which has exhibited resistance against different antibiotics in recent years. The aim of this study was to investigate the synergism effect of the silver nanoparticles with the shallot and nettle alcoholic extracts against the standard and multidrug resistant A. baumannii isolates. Samples were collected from intensive care units and the A. baumannii isolates were identified using biochemical tests. Then, the antibiogram test was carried out for each isolate. The antibacterial effect of nanoparticles, shallot and nettle extracts was evaluated singularly and in combination with each other against standard and resistant A. baumannii isolates. Measuring the diameter of inhibited growth zone, MIC, MBC and checkerboard tests were conducted for each isolate. The results showed that the silver nanoparticles, shallot and nettle alcoholic extracts each had antibacterial property against the standard and resistant A. baumannii isolates. The mixture of the nettle extract with silver nanoparticles had a synergism effect against the standard and resistant isolates and the mixture of the shallot extract with silver nanoparticles had an additive effect against A. baumannii isolates. Due to the increase of antibiotics resistance and the resistance to the pathogenic bacteria especially in intensive care units, it is necessary to find effective and accessible substances to destroy the resistant bacteria and reduce the mortality rate of patients. The results of the present study revealed that the antibacterial property of the shallot and the nettle alcoholic extracts could increase the antibacterial property of the silver nanoparticles. As a result, these can be used for disinfecting different wards of a hospital, in particular, the intensive care units.

The present study aims to investigate the role of low molecular weight compound heparin sodium salt (HSS) to control the differentiation of the human umbilical cord (UC) derived mesenchymal stem cells (MSCs) through possible interaction with WWTR1 protein. In order to carry out this study, the human UC-derived stem cells were isolated and characterized by stem cell specific markers and the effect of HSS was studied by altering the phenotypes of MSCs. An Insilco approach was employed to reveal the structural determination of the ligand, the WWTR1 protein binding site and to predict the strength of the interaction. After HSS treatment, WWTR1, Oct4, nanog, SOX9 gene expressions were studied using real-time polymerase chain reaction (PCR). Cell staining was performed using alizarin red to confirm the formation of osteocytes. Mineralization indicated by osteocytes was confirmed using alizarin red after the treatment of HSS. Post, HSS treatment, OCT4, Nanog, RUNX2, COL1A1 and WWTR1 gene expressions were positively modulated. Heparin treatment of MSCs lead to the up regulation of WWTR1 along with the down regulation of stemness markers Oct4 and Nanog expression. In silico studies also predicted the possible interaction of WWTR1 with HS. Results indicated that Amino acid residues ASP57, GLN83, GLN109, THR135, and TYR141 came up as a prominent interaction centre; ASP57, GLN83 and THR135 recorded the highest interaction energy – while ASP57 mostly participated in an electrostatic interaction. To conclude, it can be stated that heparin can possibly interact with WWTR1 along with having the capability to direct cells towards osteogenic lineages.

In many parts of the world, a rich tradition of using herbal medicine have been formed through history for treating many infectious diseases. Because of the side effects and the resistance that pathogenic microorganisms build against the antibiotics, much recent attention has been paid to extract biologically active compounds from plant species used in herbal medicine. The aim of this study was to determine the antibacterial activity of four medicinal plants’ essential oils from Labiatae family – Salvia macrosiphon, Rosmarinus officinalis, Dracocephalum polychaetum, and Origanum vulgare – on four pathogenic bacteria as well as identify their chemically active compound. The essential oil was extracted using hydro-distillation method. Then, analyzed by gas chromatography (GC) and gas chromatography-mass spectrometry (GC/MS) and compounds were identified. The antibacterial activity of the oil was evaluated using agar disc diffusion method. Minimum inhibitory concentration (MIC) and Minimum Bactericidal Concentration (MBC) were determined in a broth dilution assay. Most of the evaluated bacteria species were sensitive to the oils. MICs and MBCs showed that these 4 plants relatively had high efficacy against bacteria. More than 90% of the chemical contents of the oils was determined. The obtained results suggest and justify using the indigenous plants’ essential oils in traditional medicine as a treatment for microbial infections or as a preservative in food.

Nowadays, increasing environmental risks to human health have caused increasing attention to use chemical nitrogen sources efficiently or nitrogen supplying from organic amendments. In this respect, chicken manure seems to be a good alternative to chemical nitrogen fertilizers. The purpose of this experiment is to investigate the role of urea and poultry manure and ecological factors (climate) on the quantity of some chemical compositions of the potato tuber. To study the effects of urea and poultry manure on some potato tuber compositions, the factorial experiment was conducted in 2015-2016 during the growing season in 2 regions; Azna and Khorramabad which is located in Lorestan province in the west of Iran. The factors were 2 different sources of nitrogen supply including urea fertilizer and poultry manure. Results show that the highest activity of nitrate reductase (NR) in both sites was observed using 10304 kg of poultry manure ha-1. The results also showed that the tuber nitrate content was only affected by the fertilizer source. In both studied places, the highest tuber nitrate was associated with high levels of urea fertilizer. Based on the results, the vitamin C of potato tubers grown by poultry manure were also superior to the relative vitamin C content. According to the results of this experiment, the accumulation of more than 2 times the nitrate in the tubers was obtained from urea fertilizer (244.2 versus 100 ppm). In order to achieve food security and an efficient and sustainable food chain, chemical fertilizers must be replaced with organic fertilizers. Thus, the belief that the use of organic fertilizers, at any level, would not cause any problem for the health of products is definitely incorrect.