Rapid and sensitive detection of Mycobacterium tuberculosis based on strand displacement amplification and magnetic beads

Tuberculosis is one of the main infectious diseases threatening public health, and the development of simple, rapid, and cost‐saving methods for tuberculosis diagnosis is of profound importance for tuberculosis prevention and treatment. The bacterium Mycobacterium tuberculosis (MTB) is the pathogen that causes tuberculosis, and assaying for MTB is the only criterion for tuberculosis diagnosis. A new enzyme‐free method based on strand displacement amplification and magnetic beads was developed for simple, rapid, and cost‐saving MTB detection. Under optimum conditions, a good linear relationship could be observed between fluorescence and MTB specific DNA concentration ranging from 0.05 to 150 nM with a correlation coefficient of 0.993 (n = 8) and a detection limit of 47 pM (3σ/K). The present method also distinguished a one base mismatch from MTB specific DNA, showing great promise for MTB genome single base polymorphism analysis. MTB specific DNA content in polymerase chain reaction samples was successfully detected using the new method, and recoveries were 97.8–100.8%, indicating that the present method had high accuracy and shows good potential for the early diagnosis of tuberculosis.     

Energy response of radiophotoluminescent glass dosimeter for diagnostic kilovoltage x-ray beams

PurposeThis study aimed to investigate the energy response of a radiophotoluminescent glass dosimeter (RGD) for diagnostic kilovoltage x-ray beams by Monte Carlo (MC) calculations and measurements.MethodsThe uniformity and reproducibility of GD-352M (with Sn filter) and GD-302M (no filter) were tested with 45 RGDs in free air. Subsequently, the RGD response was obtained as a function of an Al-HVL using the parameter, quality index (QI), which is defined as the ratio of the effective energy (keV) to the maximum energy (keV) of the photons. The x-ray fluence spectra with QI of 0.4, 0.5, and 0.6 were set for tube voltages of 50 ~ 137.6 kVp. The RGD response was calculated in free air using the MC method and verified by the air kerma, K_air, measured using an ionization chamber.ResultsThe uniformity and reproducibility of the 45 RGDs were ± 2.3% and ± 2.7% for GD-352M and ± 0.7% and ± 1.6% for GD-302M at the one standard deviation level, respectively. The calculated RGD response was 0.965 to 1.062 at Al-HVL 2.73 mm or more for GD-352M and varied from 3.9 to 2.8 for GD-302M. Both RGD responses exhibited a good correlation with the Al-HVL for the given QI. K_air measured by RGDs for each beam quality with a QI of 0.5 was in the range of −5%~0.8% for GD-352M and −1.8%~3% for GD-302M, relative to the chamber measurements.ConclusionsThe RGD response was indicated as a function of the Al-HVL for the given QI, and it presented a good correlation with the Al-HVL.     

The cornerstone of integrating circulating tumor DNA into cancer management

Recent circulating tumor DNA (ctDNA) research has demonstrated its potential as a non-invasive biomarker for cancer. However, the deployment of ctDNA assays in routine clinical practice remains challenging owing to variability in analytical approaches and the assessment of clinical significance. A well-developed, analytically valid ctDNA assay is a prerequisite for integrating ctDNA into cancer management, and an appropriate analytical technology is crucial for the development of a ctDNA assay. Other determinants including pre-analytical procedures, test validation, internal quality control (IQC), and continual proficiency testing (PT) are also important for the accuracy of ctDNA assays. In the present review, we will focus on the most widely used ctDNA detection technologies and the key quality management measures used to assure the accuracy of ctDNA assays. The aim of this review is to provide useful information for technology selection during ctDNA assay development and assure a reliable test result in clinical practice.     

Weathering of Basalts by Aspergillus sp. FS-4 Strain: Glass Compositions are Prone to Weathering

Abstract

Mechanisms underlying microbe-induced dissolution of basalt and its effect on glass composition remain unclear. Two powdered basalts, with a glass composition of 27?vol% (Basalt-T) and 18?vol% (Basalt-F), respectively, were inoculated with Aspergillus sp. FS-4 in Czapek medium. Changes in pH, siderophores and main dissolved elements were examined. Basalt-F led to a greater decrease in pH and higher production of siderophores than Basalt-T mainly due to the fungi activity. Element dissolution was 2 to 3 fold higher in Basalt-T than in Basalt-F, and the dissolution was non-stoichiometric for both basalts. FS-4 appears important for basalt weathering. Composition of geologic material had a considerable impact on weathering. Materials with a higher glass percentage are prone to weathering. Further quantitative evaluation is required.     

Improved Methods for Making and Using Glass Knives

We have observed over time that the right side of a glass knife is the optimal cutting edge for microtomy if the counterpiece (heel opposite the edge) is controlled within 1 mm. The right cutting edge has been considered the “saw toothed” side and has not been used for ultrathin sectioning. We have observed that the right cutting edge is sharper and more durable than the left. Light and scanning electron microscopy were used to observe the cutting edge, and transmission electron microscopy was used to examine semithin and ultrathin sections of animal and plant tissues cut by the right and left sides of the cutting edge. The results indicate that the cutting edge becomes sharper and more durable from left to right. Both the quality and efficiency of ultrathin sectioning is improved by using the right cutting edge.     

Structural aspects of ganglioside-containing membranes

The demand for understanding the physical role of gangliosides in membranes is pressing, due to the high number of diverse and crucial biological functions in which they are involved, needing a unifying thread. To this purpose, model systems including gangliosides have been subject of extensive structural studies. Although showing different levels of complication, all models share the need for simplicity, in order to allow for physico-chemical clarity, so they keep far from the extreme complexity of the true biological systems. Nonetheless, as widely agreed, they provide a basic hint on the structural contribution specific molecules can pay to the complex aggregate. This topic we address in the present review. Gangliosides are likely to play their physical role through metamorphism, cooperativity and demixing, that is, they tend to segregate and identify regions where they can dictate and modulate the geometry and the topology of the structure, and its mechanical properties. Strong three-dimensional organisation and cooperativity are exploited to scale up the local arrangement hierarchically from the nano- to the mesoscale, influencing the overall morphology of the structure.     

Detection and Capturing of <Superscript>14</Superscript>C Radioactively-Labeled Small Subunit rRNA from Mixed Microbial Communities of a Microbial Mat Using Magnetic Beads

Carbon cycling in the hypersaline microbial mats from Chiprana Lake, Spain is primarily dependent on phototrophic microorganisms with the ability to fix CO₂ into organics that can be further utilized by aerobic as well as anaerobic heterotrophic bacteria. Here, mat pieces were incubated in seawater amended with ¹⁴C sodium bicarbonate and the incorporation of the radiocarbon in the small subunit ribosomal RNA (SSU rRNA) of mat organisms was followed using scintillation counter and autoradiography. Different domains of SSU rRNA were separated from the total RNA by means of streptavidin-coated magnetic beads and biotin-labeled oligonucleotide probes. The ¹⁴C label was detected in isolated RNA by both scintillation counter and autoradiography, however the latter technique was less sensitive. Using scintillation counter, the radiolabel incorporation increased with time with a maximum rate of 0.18 Bq ng⁻¹ detected after 25 days. The bacterial SSU rRNA could be captured using the magnetic beads, however the hybridization efficiency was around 20%. The captured RNA was radioactively labeled, which could be mainly due to the fixation of radiocarbon by phototrophic organisms. In conclusion, the incubation of microbial mats in the presence of radiolabeled bicarbonate leads to the incorporation of the ¹⁴C label into RNA molecules through photosynthesis and this label can be detected using scintillation counter. The used approach could be useful in studying the fate of fixed carbon and its uptake by other microorganisms in complex microbial mats, particularly when species-specific probes are used and the hybridization efficiency and RNA yield are further optimized.     

Alginate-entrapped Haslea ostrearia as inoculum for the greening of oysters

Entrapment in calcium alginate beads of the marine diatom, Haslea ostrearia, was successfully used for stock-culture managment and afterwards the sowing of ponds for the greening of oysters. After storage during almost 2 months, viable and cultivable cells were recovered from beads by dissolving alginate matrix but an original way lies in directly introducing beads in ponds and promoting natural cell leakage. © Rapid Science Ltd. 1998