Comparison of PCR-RFLP with 21-plex PCR and rDNA Sequencing for Identification of Clinical Yeast Isolates

Non-albicans Candida species and other rare yeasts have emerged as major opportunistic pathogens in fungal infections. Identification of opportunistic yeasts in developing countries is mainly performed by phenotypic assay, which are time-consuming and prone to errors. The aim of the present study was to evaluate PCR-RFLP as a routinely used identification technique for the most clinically important Candida species in Iran and make a comparison with a novel multiplex PCR, called 21-plex PCR. One hundred and seventy-three yeast isolates from clinical sources were selected and identified with sequence analysis of the D1/D2 domains of rDNA (LSU rDNA) sequencing as the gold standard method. The results were compared with those obtained by PCR-RFLP using MspI restriction enzyme and the 21-plex PCR. PCR-RFLP correctly identified 93.4% of common pathogenic Candida species (C. albicans, C. glabrata, C. parapsilosis, C. tropicalis, and P. kudriavsevii (=?C. krusei)) and was able to identify 45.5% of isolates of the uncommon yeast species compared to the D1/D2 rDNA sequencing. Compared with PCR-RFLP, all common Candida species and 72.7% of uncommon yeast species were correctly identified by the 21-plex PCR. The application of the 21-plex PCR assay as a non-sequence-based molecular method for the identification of common and rare yeasts can reduce turnaround time and costs for the identification of clinically important yeasts and can be applied in resource-limited settings.

     

Radiation dose and cancer risks from radiation exposure during abdominopelvic computed tomography (CT) scans: comparison of diagnostic and radiotherapy treatment planning CT scans

In the present study, radiation doses and cancer risks resulting from abdominopelvic radiotherapy planning computed tomography (RP-CT) and abdominopelvic diagnostic CT (DG-CT) examinations are compared. Two groups of patients who underwent abdominopelvic CT scans with RP-CT (n = 50) and DG-CT (n = 50) voluntarily participated in this study. The two groups of patients had approximately similar demographic features including mass, height, body mass index, sex, and age. Radiation dose parameters included CTDI_vol, dose–length product, scan length, effective tube current, and pitch factor, all taken from the CT scanner console. The ImPACT software was used to calculate the patient-specific radiation doses. The risks of cancer incidence and mortality were estimated based on the BEIR VII report of the US National Research Council. In the RP-CT group, the mean ± standard deviation of cancer incidence risk for all cancers, leukemia, and all solid cancers was 621.58 ± 214.76, 101.59 ± 27.15, and 516.60 ± 189.01 cancers per 100,000 individuals, respectively, for male patients. For female patients, the corresponding risks were 742.71 ± 292.35, 74.26 ± 20.26, and 667.03 ± 275.67 cancers per 100,000 individuals, respectively. In contrast, for DG-CT cancer incidence risks were 470.22 ± 170.07, 78.23 ± 18.22, and 390.25 ± 152.82 cancers per 100,000 individuals for male patients, while they were 638.65 ± 232.93, 62.14 ± 13.74, and 575.73 ± 221.21 cancers per 100,000 individuals for female patients. Cancer incidence and mortality risks were greater for RP-CT than for DG-CT scans. It is concluded that the various protocols of abdominopelvic CT scans, especially the RP-CT scans, should be optimized with respect to the radiation doses associated with these scans.

     

The enhanced expression of heat stress-related genes in scleractinian coral ‘Porites harrisoni’ during warm episodes as an intrinsic mechanism for adaptation in ‘the Persian Gulf’

Coral Reefs - Shallow water tropical reefs are widely threatened by anthropogenic ocean warming which sometimes exceeds their thermal tolerance limit. The majority of reefs have been currently...     

Synthesized Zn(II)-Amino Acid and -Chitosan Chelates to Increase Zn Uptake by Bean (Phaseolus vulgaris) Plants

Journal of Plant Growth Regulation - Plants require optimum amounts of nutrients for suitable growth and yield production. Accordingly, the most efficient methods of fertilization, including the...     

Coarser taxonomic resolutions are informative in revealing fish community abundance trends for the world’s warmest coral reefs

Coral Reefs - The Arabian Gulf is a natural laboratory to examine how subtropical coral reef ecosystems might change in responding to recurring heating events because of uniquely high water...     

ABA and BAP improve the accumulation of carbohydrates and alter carbon allocation in potato plants at elevated CO2

Elevated CO₂ interactions with other factors affects the plant performance. Regarding the differences between cultivars in response to CO₂ concentrations, identifying the cultivars that better respond to such conditions would maximize their potential benefits. Increasing the ability of plants to benefit more from elevated CO₂ levels alleviates the adverse effects of photoassimilate accumulation on photosynthesis and increases the productivity of plants. Despite its agronomic importance, there is no information about the interactive effects of elevated CO₂ concentration and plant growth regulators (PGRs) on potato (Solanum tuberosum L.) plants. Hence, the physiological response and source-sink relationship of potato plants (cvs. Agria and Fontane) to combined application of CO₂ levels (400 vs. 800 µmol mol⁻¹) and plant growth regulators (PGR) [6-benzylaminopurine (BAP) + Abscisic acid (ABA)] were evaluated under a controlled environment. The results revealed a variation between the potato cultivars in response to a combination of PGRs and CO₂ levels. Cultivars were different in leaf chlorophyll content; Agria had higher chlorophyll a, b, and total chlorophyll content by 23, 43, and 23%, respectively, compared with Fontane. The net photosynthetic rate was doubled at the elevated compared with the ambient CO₂. In Agria, the ratio of leaf intercellular to ambient air CO₂ concentrations [C_i:C_a] was declined in elevated-CO₂-grown plants, which indicated the stomata would become more conservative at higher CO₂ levels. On the other hand, the increased C_i:C_a in Fontane showed a stomatal acclimation to higher CO₂ concentration. The higher leaf dark respiration of the elevated CO₂-grown and BAP + ABA-treated plants was associated with a higher leaf soluble carbohydrates and starch content. Elevated CO₂ and BAP + ABA shifted the dry matter partitioning to the belowground more than the above-media organs. The lower leaf soluble carbohydrate content and greater tuber yield in Fontane might indicate a more efficient photoassimilate translocation than Agria. The results highlighted positive synergic effects of the combined BAP + ABA and elevated CO₂ on tuber yield and productivity of the potato plants.     

Silver-coated magnetic nanoparticles as an efficient delivery system for the antibiotics trimethoprim and sulfamethoxazole against E. Coli and S. aureus: release kinetics and antimicrobial activity

BioMetals - Trimethoprim and sulfamethoxazole are prescribed for a broad spectrum of bacteria. However, the use of these medicines is restricted due to the risk of microbial resistance in the body....     

Stable polymer bilayers for protein channel recordings at high guanidinium chloride concentrations

The use of chaotropic reagents is common in biophysical characterization of biomolecules. When the study involves transmembrane protein channels, the stability of the protein channel and supporting bilayer membrane must be considered. In this letter, we show that planar bilayers composed of poly(1,2-butadiene)-b-poly(ethylene oxide) diblock copolymer are stable and leak-free at high guanidinium chloride concentrations, in contrast to diphytanoyl phosphatidylcholine bilayers, which exhibit deleterious leakage under similar conditions. Furthermore, insertion and functional analysis of channels such as α-hemolysin and MspA are straightforward in these polymer membranes. Finally, we demonstrate that α-hemolysin channels maintain their structural integrity at 2 M guanidinium chloride concentrations using blunt DNA hairpins as molecular reporters.