Journal of Experimental and Molecular Biology
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ENHANCING POTATO VIRUS X DIAGNOSIS: CHELEX™ 100 RESIN RNA EXTRACTION METHOD EVALUATION
Get PDFPotato Virus X (PVX) is a prevalent pathogen affecting potato crops, leading to substantial yield losses. This study investigates the efficacy of Chelex™ 100 resin RNA extraction method for PVX detection, through RT-qPCR analysis. We evaluate RNA yield, sensitivity, and detection limits of this method compared with Phenol-chloroform RNA extraction protocol. Our results demonstrate that Chelex™ 100 resin extraction yields superior RNA concentrations and exhibits greater sensitivity compared to ELISA immunoassay diagnostic test. RT-qPCR successfully detects PVX RNA in dilutions up to 1:100 using Chelex™ 100 resin extraction. These findings highlight the potential of Chelex™ 100 resin extraction coupled with RT-qPCR as a rapid and reliable method for PVX detection
Anti-Candida activity of flavonoids - an overview
Get PDFFlavonoids are a group of plant polyphenols which received an increased attention during the recent past due to their important antimicrobial activities. Those compounds could be a reliable source of new antifungals, used to efficiently control infections caused by pathogenic fungi such as Candida spp. Candida species represents a leading cause of mortality all around the world, posing a serious threat to medical systems. Therefore, finding new compounds with antifungal activity for treatment of Candida infections is a real challenge of modern medicine. This review focuses on the antifungal activity of natural, semi-synthetic and synthetic flavonoids against the most prevalent pathogenic Candida species. In addition, the review outlines the mechanisms of action and the possible use of flavonoids as anti-virulence agents to withstand Candida pathogenicity and antifungal resistance
MITOCHONDRIA : AS A PROTAGONIST IN NEUROLOGICAL DISORDERS IN BRIEF OVERVIEW: Role of mitochondria in neurological disorders
No full textNeurological disorders pose a great burden in general health. It is not astounding that mitochondrial malfunction emerging as a leading factor in myriad of neurological disorders. Mitochondria are extremely active cell organelles performing various functions most importantly, providing ATP to sustain cellular processes. Mitochondrial dysfunction results in altered neuronal bioenergetics, redox equilibrium and dynamics of cell and acts as focal point of pathogenesis in many human diseases including neurological disorders. Mitochondrial dynamics regulates pathways involving oxidative stress and apoptosis. Often mitochondrial division imbalance and fusion leads to mitochondrial functional impairment. Extreme variations in mitochondrial fusion causes increased mutation rate which along with increased oxidative stress can facilitate development of various neurological disorders such as Parkinson’s disease, Alzheimer’s disease, Huntington’s diseases and so on. Mitochondria has a key role in regulation of apoptosis. Mitochondrial dysfunction and mutations can have deleterious effects on neuronal functioning as neurons have high energy demand with restricted regenerative potential. Certain neuroprotective agents restores the functions of mitochondria and acts therapeutic regimens of neurodegenerative diseases
The molecular basis of the Fragile X syndrome
Get PDFThis analysis aimed to clarify the molecular basis of fragile X syndrome and explain the role of genetic material in the genetic disease's development and treatment. Fragile X syndrome is an X-linked mutation inheritance disorder. The mutated gene is called FMR-1. This is important for normal brain development and synaptic plasticity, which was verified and recognized in 1991, and this has become a hope for more clarification. FMR1 influences the translation of messenger RNA (mRNA), but identifying functional targets was complex and directly related to translational control and showed that dysregulated translation initiation signaling was observed for the FMR1 gene in the FMR1 knockout mouse model of FXS.
Because of the epigenetic alteration, such as hypermethylated at the DNA promoter region, and chromatin modification, such as H3K9 methylation, the FMR1 gene can be imprinted. Still, their mechanisms of aberrant epigenetic marks play a role in the etiology of many neurodevelopmental disorders, some of which we still do not fully understand and need to show more. The opportunities for epigenetic markers to map and alter epigenetic marks and the potential for therapies based on epigenetics and noncoding manipulation. For neurodevelopmental and behavioral conditions, including mental retardation, autism, anxiety, and mood disturbance, FMR1 loss of control is a model.
Most studies have focused on the new and effective approach for Fragile X syndrome, which is Gene therapy is unarguably the definitive way to treat and possibly cure genetic diseases. Many of them are under clinical trial, but more studies, such as the CRISPR/Cas9-based method, should be approved. Adeno-associated viral (AAV) vectors are highly effective for generating models. Most research is used in the mouse model of fragile X syndrome, where AAVs have been used to express fragile X mental retardation protein (FMRP), which is missing or highly reduced in the disorder. The vast expansions need southern blotting in myotonic dystrophy. Fragile X is diagnosed by a form of Southern blotting that relies on the size and the FRAXA gene's methylation status. Almost always, genetic testing is performed by PCR. The few Southern blotting uses include checking for significant destructive gene rearrangements and complete mutations of fragile X and myotonic dystrophy. Expansions suppress the expression of closely adjacent genes, causing loss of function. A named FMRI (fragile-X mental retardation syndrome) gene cDNA probe. Some unique molecular mechanisms, such as CGG expansion in Fragile-X syndrome, can make a particular sequence change in a gene far more probable than any other change
From Erlenmeyer flask to microplates – lessons learned from measuring Escherichia coli growth
Get PDFGrowth curve measurements are fundamental operations in microbiology and molecular biology. Microplate readers have the advantage of high-throughput and automation. This allows for real-time monitoring of many bacterial cultures and a decrease in labor and costs. The volume and geometry of a well from a microplate imply that calibration and comparative studies must be performed before switching from one cultivation and growth monitoring system to another. Here, Escherichia coli XL1 Blue was used as a reference strain in order to test the feasibility of using a common, filter-based microplate reader for recording growth curves when performing antibiotic and toxicity testing. 
COLD ATMOSPHERIC PLASMA TREATMENT MODULATES THE EXPRESSION OF cdk1, tnfα AND tp53 GENES IN HUMAN OSTEOSARCOMA CELLS
Get PDFOsteosarcoma (OS), a malignancy primarily affecting children and adolescents, is the most frequently encountered malignant, non-hematologic, bone tumor. Despite the gradual improvement of survival rates, the management of this disease remains problematic due to challenges such metastasis development, its heterogeneous characteristics, and resistance to cytostatic drugs. Cold atmospheric plasma (CAP), a partially ionized gas operating at near room temperature, which is comprised of free carriers, excited or neutral molecules, and active radicals capable of initiating diverse physical phenomena and chemical reactions represent a new and innovative potential solution in cancer therapy. The aim of this study was to evaluate the capacity of CAP produced by our custom-build plasma source to induce cytotoxic effects to HOS cells and to analyze post-treatment modulations of cdk1, tnfα and tp53 genes expression. Direct and indirect CAP treatments effectiveness on HOS cells were evaluated by MTT assay and the regulation of interest genes expression were carried out by RT-qPCR analysis. Cell viability analysis revealed a strong cytotoxic effect of direct CAP treatment on HOS cells, while the indirect treatment resulted in a slight decrease of cells viability. Direct CAP treatment modulates the expression of all analyzed genes, both at 2- and 24-hours post-treatment. In conclusion, direct CAP treatment produced by our custom-build plasma source have cytotoxic effects on HOS cells in a dose-dependent manner up to 24 hours post-treatment. Furthermore, direct CAP treatment induces cell cycle arrest of HOS cells, and the CAP-induced cell death is independent of tp53 gene
EVALUATION OF THE NEUROPROTECTIVE EFFECTS OF HYDROETHANOLIC EXTRACT OF DIOSPYROS MESPILLIFORMIS TRUNK BARK (EBENACEA) ON DIAZEPAM-INDUCED AMNESIA IN MICE
Get PDFDiospyros mespilliformis (DM) is a plant of the Ebonaceae family that is commonly used in traditional medicine to treat epilepsy and schizophrenia. Radial arm mazes (RAM), Y mazes, and the novel object recognition test (NOR) were used to assess working and reference memory in mice. After the different tests, the hippocampus of the animals was separated for the determination of biochemical markers such as acetylcholinesterase (AchE), malondialdehyde (MDA), and superoxide dismutase (SOD). The plant extract produced a significant rise in the percentage of alternation of mice in the Y-maze test, the time to observe the new object at the dose of 50 mg/kg (p<0.01), and the discrimination index (p<0.001) in all animals treated with the extract compared to the negative control. The result of the RAM test showed a significant decrease (p<0.05) in the number of errors on the reference memory in animals treated with the extract at 100 mg/kg. The extract significantly decreased AchE activity, and MDA concentration, and significantly increased SOD activity compared to the negative control group. These effects of the hydroethanolic extract on working memory and reference DM extract would be related to the phenolic compounds quantified in the plant extract
Distribution of the 23-bp polymorphism of the prion protein gene in Jersey cattles in Turkey
No full textBackground: Bovine spongiform encephalopathy (BSE) is a prion disease that is always fatal in cattle and is considered an important risk factor for human health. Genetic polymorphisms that alter prion proteins may be associated with susceptibility or resistance to infectious spongiform encephalopathy. Therefore, we investigated the distribution of the 23 bp indel variant in the prion protein (PRNP) gene in Jersey cattle in Turkey.
Methods: A total of 95 unrelated Jersey cattle (79 of reproductive age and 16 of non-reproductive age) from a private farm in Izmir were included in the study. Genomic DNA was obtained from the milk of reproductive-age cattle and the saliva of non-reproductive-age cattle. A 23-bp indel polymorphism in the PRNP gene promoter region was genotyped by polymerase chain reaction (PCR) analysis.
Results: The three genotypes of the PRNP 23-bp indel variant were classified as follows: (223 bp), I/D (both 223 and 200 bp fragments), and D/D (200 bp). The frequencies of the I/I, I/D, and D/D genotypes of the PRNP 23-bp indel variant in cattle were 22 (23.16%), 48 (50.53%), and 25 (26.32%). We then examined genotype and allele distribution according to service period. No significant difference was detected in terms of PRNP gene 23 bp-indel variant genotype and allele distribution in the groups created according to the service period (p>0.05).
Conclusion: Although the PRNP gene 23 bp-indel variant genotype and allele distribution in jersey-type cattle in Turkey did not differ according to service period, our results may benefit the understanding of the genetic characteristics of the PRNP gene in cattle breeds
YEW EXTRACTS - A POSSIBLE SOURCE OF BIOACTIVE COMPOUNDS WITH POTENTIAL ALLELOPATHIC PROPERTIES
Get PDFRepresentative species of the Taxus genus, known since antiquity for their toxicity, have been an essential plant resource for the production of plant extracts with medicinal properties (anti-inflammatory, antifungal, and antibacterial) and, more recently, for the study of their allelopathic properties with significant implications in organic agriculture (bioherbicidal potential). With a view to the development of this new direction of research, the present work aims: to present a qualitative evaluation of two types of plant extracts (aqueous and alcoholic) of different concentrations (1% and 5%) obtained from various organs (bark, leaves, arils, and seeds) belonging to three Taxus taxa: a spontaneous taxa - Taxus baccata L. (T1) and two cultivated taxa - Taxus baccata (T2) and Taxus baccata 'Robusta' (T3) at different times of the phenological cycle by determining their absorption spectra (of both types of extracts), total amounts of polyphenols and flavonoids, and by evaluating their antioxidant capacity (alcoholic extracts); to investigate, under experimental conditions of cultivation the possible allelopathic effects induced by the aqueous extracts on the germination and development of the seedlings in two test plant species: Amaranthus retroflexus L. (ruderal species) and Lycopersicon esculentum Mill, variety Silvia (crop species). The data indicates the presence of phenolic and compounds, alkaloids, and carotenoid pigments in the alcoholic extracts prepared from the different organs of the studied yew taxa, showing higher amounts of polyphenols and flavonoids in the extracts obtained from the leaves of taxon T2 compared to taxon T1. The effect of 1% and 5% aqueous extracts obtained from the bark of the three investigated yew taxa, as well as 1% aqueous extracts prepared from the leaves of taxon T3 on seed germination and seedling growth was more pronounced in the crop species Lycopersicon esculentum Mill, variety Silvia, compared to the ruderal broadleaf weed Amaranthus retroflexus L., both species having rapid germination stimulated by ambient light. Aqueous extracts of 5% concentration obtained from the arils of plants belonging to the three yew taxa stimulated the elongation of the seedlings’ hypocotyls of both test species
The epigenetic of the panic disorder
Get PDFFrom this review, I discussed the epigenetics of panic disorder .epigenetic is the changes in the heritable phenotype without any change from the DNA sequence. The roles of different types of epigenetics (DNA methylation and chromatin remodeling) have been controversial from one study to another. The researcher focused on the roles of the candidate genes in the development and progress of panic disorder .there different candidate genes located the different chromosomes such as 1q, 2p, 2q, 3, 7, 9, 11, 12q13, 12q23, and 15 that may be used as markers give for the future promise to diagnosis panic disorder in early life span. Recently, no studies have demonstrated gene therapy's roles in treating panic disorder, and the basic contribution of interventions such as exposure, cognitive therapy, relaxation training, and breathing retraining has not yet been identified. Differences between these patients that assess the efficacy of exposure-based cognitive-behavioral therapy and related neuroplastic changes can be demonstrated by altered protection signal processing and anterior cingulate cortex-amygdala couplin