A Robust DNA Amplification Fingerprinting System Applied to Analysis of Genetic Variation Within Fusarium oxysporum f.sp. cubense

Genetic variation among isolates of F. oxysporum f.sp. cubense (Foc) was analysed using a DNA amplification fingerprinting (DAF) system modified to improve reproducibility and transportability. This analysis was done after determining the widest tolerance range (or 'window of reproducibility') for each component in amplification reaction. Reproducible polymerase chain reactions (PCRs) were achieved with between 25 and 250 ng of template DNA, 9–15 μM primer, 4–6 mM MgCl² and 2–4 units of Stoffel Fragment enzyme. For experimental work we used the middle value of these ranges which allowed at least 20% error tolerance for each component. Similarly, thermocycling and electrophoresis conditions were also improved. Manual scoring of the DNA fingerprints was compared to analysis of scanned gel images using the Gel Compar program (Applied Maths, Kortrijk, Belgium). The data were clustered by unweighted pair group method analysis (UPGMA) based on the Jaccard similarity coefficient. Isolates of Foc representing all known vegetative compatibility groups (VCGs) were examined and the genetic relationships between the VCGs were determined. Isolates of Foc were divided into two major groups with 30% genetic similarity. These optimized DNA amplification, thermocycling, and electrophoresis conditions were suitable for analysis of other organisms and should be applicable to other techniques that use arbitrary primers such as random amplified polymorphic DNA (RAPD) and arbitrarily primed-PCR (AP-PCR).     

An eco-friendly matrix-augmented fluorescence spectroscopic approach for the analysis of mitoxantrone,an oncogenic therapy; application to the dosage form and biological matrices

Mitoxantrone (MXN) is a synthetic anthracenedione oncogenic therapy. It is often prescribed as an anticancer agent to manage a variety of cancers. A green, fast, and easy fluorimetric technique for the assay of MXN as a topoisomerase type II enzyme suppressor. An investigation of MXN's fluorescence behavior in various media and solvents constituted the basis for this new technique. Methanol was shown to enhance the intrinsic fluorescence considerably. After excitation at 610 nm, the highest fluorescence intensity was found at 675 nm. Various experimental parameters, such as media, solvents, and pH levels, were tested and adjusted. ICH (International Conference on Harmonization) guidelines were followed when validating procedures. It was possible to achieve linearity in the 0.02–1.50 μg ml⁻¹ with the method. The sensitivity (in terms of limit of detection and limit of quantification) was 0.003 and 0.008 μg ml⁻¹, indicating low toxicity. As a result, the current technology has a remarkable recovery for detecting residues in diverse bodily fluids. Also, the quantum yield was estimated for the designed system. Finally, the method was rated by eco-scale scoring.     

The correlation of combined OGG1, CYP1A1 and GSTP1 gene variants and risk of lung cancer of male Iraqi waterpipe tobacco smokers

Molecular Biology Reports - Genetic polymorphisms of genes whose products are responsible for activities, such as xenobiotic metabolism, mutagen detoxification and DNA-repair, have been predicted...     

Betaine Significantly Improves Multiplex Tetra-Primer ARMS-PCR Methods

The tetra-primer ARMS-PCR method offers significant advantages over the commonly used methods to genotype single nucleotide polymorphisms. It offers fast and cost-effective detection and requires minimum level of expertise and basic instrumentation. The benefits of TP-ARMS-PCR increase exponentially upon multiplexing. However, several complications preclude the common use of multiplex TP-ARMS-PCR methods, primarily the lack of robustness and the difficulty of optimization. We have previously developed triplex and quadruplex TP-ARMS-PCR methods involving the simultaneous detection of up to three SNPs in a single reaction and utilized Betaine, a PCR additive used to enable amplification of GC-rich templates with strong secondary structures, in an attempt to facilitate method development and optimization. In the present communication, we introduced experimental data demonstrating the important effects of Betaine on our previous methods and its potential to overcome the ruggedness and robustness issues commonly found in TP-ARMS-PCR methods, and highlighted the general benefits of Betaine with respect to TP-ARMS-PCR. Our data support the routine inclusion of Betaine in all TP-ARMS-PCR methods, especially when multiplexing is concerned.     

Characterization and distribution of <Emphasis Type="Italic">Daucus</Emphasis> species in Syria

Syria is considered as one of the important centres for Daucus diversity including Daucus carota. Therefore, it is essential to study the distribution and characterization of these species in Syria. An exploration of plants belonging to the Apiaceae family was conducted on road and field sides in several areas of Syria. Seeds (fruits) from these plants have been also collected. The seeds were sown in pots containing peatmoss in a glasshouse and emerging plants were grown until flowering and seed formation. The plants were classified based on leaf, umbel, and seed shape. Proteins were extracted from the leaves and analysed using electrophoresis to establish genetic relationships among species. Seven Daucus species have been identified to grow in Syria. These are D. aureus, D. bicolor, D. carota, D. durieua, D. guttatus, D. littoralis, and D. muricatus. Isozyme and total protein analysis, cluster analysis, and correlation matrix have revealed considerable genetic variation among studied Daucus species. Wild carrot (D. carota) came in one group with its cultivated form (D. carota ssp. sativus) and the closest species to them was D. guttatus. Species D. bicolor and D. durieua were in the same group and D. aureus and D. littoralis were in another group farther from the previous groups. The farthest species on the genetic tree was D. muricatus.     

Glucose control of staphylococcal enterotoxin A synthesis and location is mediated by cyclic AMP

The regulation of staphylococcal enterotoxin A (SEA) synthesis in a defined medium was studied using continuous culture techniques. SEA production was repressed by glucose and repression could be overcome by addition of exogenous cyclic AMP. As well as this classical catabolite repression control, addition of glucose to de-repressed steady-state cultures resulted in rapid disappearance of toxin from the medium (also mediated by loss of cyclic AMP). When the toxin dissappeared from the medium, it was taken up again by the bacteria without apparent modification.     

Hematological indices and oxidative stress biomarkers response to the starvation of Clarias gariepinus

Starvation effects for five weeks on energy reserves, oxidative stress and hematological indices in Nile catfish Clarias gariepinus was studied. The low protein level in starved fish may result from the lowering effect of prolonged starvation on protein synthesis rather than due to its degenerating protein. Moreover, the elevated level of serum amino acids may promote gluconeogenesis in liver. In addition, the lipid depletion in starved fish may be related to the preferential uses of lipids as an energy to starve fish. Also, unchanged glycemic level may introduce a potent evidence for the presence of active gluconeogenesis, depending on both amino and fatty acids precursors. Also, kidney and liver showed disturbances in metabolites associated with oxidative damage such as elevations in total peroxide, carbonyl protein and DNA fragmentation; these may cause dysfunction to these organs after five weeks of starvation. Total peroxide, carbonyl protein and DNA fragmentation were significantly increased in gills, liver and kidney by 29.9, 30.9 and 30.5; 83.6, 84.6 and 53.7; 82.4, 43.3 and 75.7%, respectively. Starvation induced severe anemia and loss of body weight in the fish. However, white muscle did not show any oxidative damage after five weeks of starvation.     

Integrated Use of Organic and Bio-fertilizers to Improve Yield and Fruit Quality of Olives Grown in Low Fertility Sandy Soil in an Arid Environment

Olive productivity should be improved through stimulating nutrition, particularly under poor fertility soils. Consequently, the objective of this study was to assess the efficacy of applying organic and bio-fertilizers on the physiological growth, yield and fruit quality of olive trees under newly reclaimed poor-fertility sandy soil in an arid environment. During a field experiment carried out at El-Qantara, North Sinai, Egypt over two consecutive seasons (2019–2020 and 2020–2021), olive Kalamata trees were evaluated under three organic fertilizer treatments alone or in combination with three bio-fertilizers treatments. Organic fertilizer was applied as goat manure (16.8 kg/tree/year), or olive pomace (8.5 kg/tree/year) in mid-December of each season vs. untreated trees. The bio-fertilizers were applied as N-fixing bacteria (150 g/tree) was inculated in early March of each season, or amino acid mixture (1.5%) was applied three times, at 70% of full bloom, 21 days after full bloom, and a month later in comparison to a non-fertilized trees (control). The cultivar used was Kalamata, a dual-purpose cultivar for oil and table olives whose value increases when processed as table olives. The results indicated that the goat manure followed by olive pomace significantly enhanced photosynthetic pigments (chlorophyll a, b, and carotenoids), leaf mineral contents (N, P, K, Ca, Mg and Fe), tree canopy volume, number of flowers per inflorescence, number of inflorescences per shoot, initial fruit set, fruit retention. For fruit quality, fruit length and width, fruit weight, and total fruit yield was increased compared to the non-fertilized control. Likewise, The bio-fertilizer N-fixing bacteria followed by the amino acid mixture significantly improved all of the aforementioned parameters. Accordingly, it is recommended, both environmentally and economically to utilize organic and bio-fertizers, particularly goat manure combined with N-fixing bacteria, in low-fertility soil to sustain olive production as well as reducing mineral fertilization.