Current Biotechnology - Volume 9, Issue 3, 2020

Finger millet is a superior staple food for human beings. Microsatellite or Simple Sequence Repeat (SSR) marker is a powerful tool for genetic mapping, diversity analysis and plant breeding. In finger millet, microsatellites show a higher level of polymorphism than other molecular marker systems. The identification and development of microsatellite markers are extremely expensive and time-consuming. Only less than 50% of SSR markers have been developed from microsatellite sequences for finger millet. Therefore, it is important to transfer SSR markers developed for related species/genus to finger millet. Cross-genome transferability is the easiest and cheapest method to develop SSR markers. Many comparative mapping studies using microsatellite markers clearly revealed the presence of synteny within the genomes of closely related species/ genus. Sufficient homology exists among several crop plant genomes in the sequences flanking the SSR loci. Thus, the SSR markers are beneficial to amplify the target regions in the finger millet genome. Many SSR markers were used for the analysis of cross-genome amplification in various plants such as Setaria italica, Pennisetum glaucum, Oryza sativa, Triticum aestivum, Zea mays and Hordeum vulgare. However, there is very little information available about cross-genome amplification of these markers in finger millet. The only limited report is available for the utilization of cross-genome amplified microsatellite markers in genetic analysis, gene mapping and other applications in finger millet. This review highlights the importance and implication of microsatellite markers such as genomic SSR (gSSR) and Expressed Sequence Tag (EST)-SSR in cross-genome analysis in finger millet. Nowadays, crop improvement has been one of the major priority areas of research in agriculture. The genome assisted breeding and genetic engineering plays a very crucial role in enhancing crop productivity. The rapid advance in molecular marker technology is helpful for crop improvement. Therefore, this review will be very helpful to the researchers for understanding the importance and implication of SSR markers in closely related species.

The study of the interactions between enzymes and inactivators can often be performed with the help of the reporter substrate method in which the time-dependent decrease of the rate of substrate disappearance (or product formation) is monitored. In the present contribution, we wish to describe examples of the utilization of this rapid and efficient method for reactions whose rates can be monitored by spectrophotometric or fluorimetric measurements. After the collection of the data in an Excel file, a very simple program can be applied to extract the values of pseudo-- first-order rate constants. The inactivation can be complete or result in a steady-state if the inactivated adduct is not totally stable or if the inactivation reaction is reversible. Similarly, the method can be used in the cases of so-called “slow binding” inhibitors. The same type of analysis allows the easy determination of kcat/Km values for substrates for which the Km value is rather low. We show that this very rapid method (less than 5 min) yields very good values of the desired kinetic parameter even if the total absorbance variations are very low (0.1 or less). In conclusion, the described experimental approach is particularly useful when applied to the reporter substrate method but it also allows the estimation of the kcat/Km parameter even if the Km value is rather low. The authors wish to dedicate this paper to the memory of the late Michel Rinné (1941-2009) whose contribution to making the data analysis program very user-friendly was invaluable.

Background: The diseases in the heart and blood vessels such as heart attack, Coronary Artery Disease, Myocardial Infarction (MI), High Blood Pressure, and Obesity, are generally referred to as Cardiovascular Diseases (CVD). The risk factors of CVD include gender, age, cholesterol/ LDL, family history, hypertension, smoking, and genetic and environmental factors. Genome- Wide Association Studies (GWAS) focus on identifying the genetic interactions and genetic architectures of CVD. Objective: Genetic interactions or Epistasis infer the interactions between two or more genes where one gene masks the traits of another gene and increases the susceptibility of CVD. To identify the Epistasis relationship through biological or laboratory methods needs an enormous workforce and more cost. Hence, this paper presents the review of various statistical and Machine learning approaches so far proposed to detect genetic interaction effects for the identification of various Cardiovascular diseases such as Coronary Artery Disease (CAD), MI, Hypertension, HDL and Lipid phenotypes data, and Body Mass Index dataset. Conclusion: This study reveals that various computational models identified the candidate genes such as AGT, PAI-1, ACE, PTPN22, MTHR, FAM107B, ZNF107, PON1, PON2, GTF2E1, ADGRB3, and FTO, which play a major role in genetic interactions for the causes of CVDs. The benefits, limitations, and issues of the various computational techniques for the evolution of epistasis responsible for cardiovascular diseases are exhibited.

Background: Aeromonas hydrophila is a heterotrophic, Gram-negative, rod-shaped, facultative anaerobic, non-spore forming bacteria that are autochthonous and widely dispersed in marine environments. The study aims at investigating the screening of Aeromonas hydrophila from spoiled vegetables and the sediment sample collected from three different estuaries located in the Bay of Bengal (Vedharanyam, Parangipettai and Pichavaram, Tamilnadu, India) for the presence of enzymes and antimicrobial activities. Objective: Isolation, enzyme screening, antimicrobial activity of Aeromonas hydrophila from spoiled vegetables and three different estuarine microbial sediment samples for the purpose of biochemical and enzymatic analysis. Methods: The bioactive compound produced by this strain was purified by using thin-layer chromatography. Results: The purified isolate of Aeromonas hydrophila strain produces good antimicrobial activity against Aspergillus niger, Candida albicans, Staphylococcus, Klebsiella and pseudomonas species. Conclusion: These isolates producing amylase, protease, lipase, and gelatinase enzymes, which are commercially very important and used in many industries and other biochemical sectors.

Objective: The inoculants, mainly nitrogen fixers and phosphate solubilizing microbes, have an influence on plant growth attributes. The current study was conducted to assess the fertilizing activity of A. chroocaccum and B. megaterium strains on growth, yield parameters and nutrient uptake of Sorghum bicolor. Methods: The isolation and identification of the nitrogen-fixing bacterium- A. chroocaccum and phosphate solubilizing microbe- B. megaterium and the growth and yield parameters of Sorghum bicolor as well as NPK uptake levels were studied. Results: A. chroocaccum and B. megaterium exhibited the proliferation of microbial population in soil by synergistic interaction with plants, and the application increased the availability of NPK in the soil after harvest of the crop. These beneficial inoculants are also known to help in the uptake of some other nutrients. The maximum beneficial aspects of the plant were noticed in the plants inoculated with 100% Recommended Dose of Fertilizer (RDF)+ A. chroocaccum+ B. megaterium, and the least was noticed in the control plants at all growth stages and at the time of harvest. Conclusion: From this study, it can be concluded that the beneficial effect of A. chroocaccum and B. megaterium is observed in sorghum crop with improved yield and nutrient uptake. Therefore, this species can be used extensively for future inoculation of the sorghum crop for better growth and development and for good returns in an ecological way.

Background: The Klf6 gene, belonging to Krüppel-like family of C2H2 zinc finger transcription factors, is strongly associated with tumor formation through high somatic mutations in carcinomas of the prostate, liver, colon, stomach, lung, neck, pituitary gland and nervous system. Recently, Klf6 super-enhancer which strongly regulates Klf6 gene expression has been identified, and the function of Klf6 super-enhancer which regulates cell growth was studied. Objective: The development of inhibitors targeting BRD4-binding super-enhancers is a potential target therapeutic strategy for tumor therapy. However, the suppression of Klf6 super-enhancer function by BRD4 inhibitors is not known. Methods: CRISPR-Cas9 editing technique was used for the Klf6 super-enhancer deletion experiment, and the expression levels of several genes for cell clones were detected by qRT-PCR analysis and Western blotting. Cell proliferation assay was applied to evaluate the functional role of Klf6 super-enhancer using several BRD4 inhibitors. The interaction of several BRD4 inhibitors against the target protein was analyzed by molecular docking simulation. Results: JQ-1, a human BRD4 inhibitor, inhibited Klf6 gene expression and its activity in HepG2 cells in a time and dose-dependent manner while simultaneously inhibiting cell growth. Besides, BETd-246, a human BRD4 inhibitor, strongly inhibited Klf6 gene expression, significantly inhibited cell growth, and exhibited higher efficacy than JQ-1. Molecular docking studies revealed that some key residues were critical for ligand-receptor interactions by forming hydrogen bonds with ligands (JQ-1: ASN140, BETd-246: ASN140, TYR106, LYS65, GLN58, MET105, and MET53). Conclusion: Our findings suggest that KLF6 is regulated by Klf6 super-enhancer and the targeting of Klf6 super-enhancer by BRD4 inhibitors may be an effective therapeutic strategy for liver cancer therapy.

Background: Laccases are multi-copper enzymes that oxidize phenolic/aromatic compounds and represent a promising alternative to environmental decontamination processes and biotechnological applications. Objective: The effects of pH and temperature on the growth and the production of laccases during the cultivation of Pleurotus sajor-caju PS-2001 in stirred-tank bioreactor were assessed. Methods: Assays were performed at fixed pH values from 4.5 to 7.5 (28°C) and at temperatures from 24 to 36°C (pH 6.5). Results: In pH testing, larger biomass concentration (4.5 g L-1) was reached at pH 5.5, whereas concentrations of 3.7, 3.1 and 1.7 g L-1 were measured at pH 4.5, 6.5 and 7.5, respectively. With ABTS as substrate, peaks of laccases activity of 50, 30 and 24 U mL-1, at pH 6.5, 5.5 and 7.5, respectively, were detected. Under different temperatures, higher mycelial concentrations (3.0 g L-1) were quantified at 66 hours at 28°C, while concentrations below 2.0 g L-1 were observed at 24, 32, and 36°C. Maximum laccases activities of 50, 42, 6 and 5 U mL-1 were obtained at 28, 32, 24, and 36°C, respectively. In all tests, the presence of other phenol oxidases – total peroxidase, manganese peroxidase, lignin peroxidase and veratryl alcohol oxidase – was observed. Conclusion: The results indicate that variations in pH and temperature during fungal cultivation strongly affect the enzymatic activity and growth kinetics of P. sajor-caju PS-2001 in a stirredtank bioreactor.

Background: Diabetic foot ulcers (DFUs) exhibit 80% of prevalence in Mexico. Efficient tissue regeneration therapies are the key factors to avoid amputations. Objective: In this study, the healing properties of a Cu/TiO2-SiO2 nanobiocatalyst applied in DFUs were analyzed. Furthermore, the morphology and crystalline structures were characterized. Methods: The nanobiocatalyst was synthesized by a sol-gel patented method proposed by López et al. The compound was characterized by scanning and transmission electron microscopies and Xray diffraction. The nanoparticles were embedded in a polymeric gel matrix (nanogel), which was utilized as a conservative therapy for chronic non-healing DFU in 8 patients with several comorbidities and chronic complications of diabetes. Wound debridement was performed prior to the nanogel administration. The nanogel was applied over the ulcers on alternate days for different periods of time, depending on the case. Results: Significant improvement in terms of enhanced epithelialization was observed in the wound healing process after a few applications. Infection spread was limited, and tissue regeneration was enhanced, with significant healing of the ulcers observed in each case. Furthermore, the successful outcome allowed to avoid the amputations that were proposed to some of the patients. Conclusion: The study proved the efficiency of the nanobiocatalyst as a safe, conservative therapy for chronic non-healing DFUs. Further investigation must be carried out to fully elucidate the wound-healing mechanisms of the nanoparticles.