Bacterial Cell Inactivation Using a Single-Frequency Batch-Type Ultrasound Device

Ultrasound technology employs cavitation to generate high-pressure soundwaves to disrupt bacterial cells. This study reveals the effectiveness of a single frequency ultrasound device for bacterial cell inactivation. A low-cost ultrasound device having a single frequency, i.e. 22 kHz for lab-scale application, was developed first, and the prototype was mechanically designed and analyzed using the finite-element method to assure the targeted natural frequency could be achieved. The prototype was then tested inactivating bacterial cells, Escherichia coli (E. coli) and Bacillus subtilis (B. subtilis), in a simple medium and a food system, and the results were then compared to a commercial system. A treatment time of up to 15 minutes was able to reduce E. coli and B. subtilis cells by 3.3 log and 2.8 log, respectively, and these results were similar to those of the commercial system. The effectiveness of bacterial cell inactivation using the developed single-frequency ultrasound device is then discussed. The findings are useful for designing low-cost ultrasound devices for application in the food industry.

Analysis of the effects of Cu-MOFs on fungal cell inactivation

Cu-MOFs containing glutarates and bipyridyl ligands produce antifungal effects on C. albicans cells and A. niger spores, and induce apoptosis-like death of the fungi, which was probably caused by the elevated level of intracellular reactive species.

Identification of a glycan cluster in gp120 essential for irreversible HIV-1 lytic inactivation by a lectin-based recombinantly engineered protein conjugate

We previously discovered a class of recombinant lectin conjugates, denoted lectin DLIs (‘dual-acting lytic inhibitors’) that bind to the HIV-1 envelope (Env) protein trimer and cause both lytic inactivation of HIV-1 virions and cytotoxicity of Env-expressing cells. To facilitate mechanistic investigation of DLI function, we derived the simplified prototype microvirin (MVN)-DLI, containing an MVN domain that binds high-mannose glycans in Env, connected to a DKWASLWNW sequence (denoted ‘Trp3’) derived from the membrane-associated region of gp41. The relatively much stronger affinity of the lectin component than Trp3 argues that the lectin functions to capture Env to enable Trp3 engagement and consequent Env membrane disruption and virolysis. The relatively simplified engagement pattern of MVN with Env opened up the opportunity, pursued here, to use recombinant glycan knockout gp120 variants to identify the precise Env binding site for MVN that drives DLI engagement and lysis. Using mutagenesis combined with a series of biophysical and virological experiments, we identified a restricted set of residues, N262, N332 and N448, all localized in a cluster on the outer domain of gp120, as the essential epitope for MVN binding. By generating these mutations in the corresponding HIV-1 virus, we established that the engagement of this glycan cluster with the lectin domain of MVN*-DLI is the trigger for DLI-derived virus and cell inactivation. Beyond defining the initial encounter step for lytic inactivation, this study provides a guide to further elucidate DLI mechanism, including the stoichiometry of Env trimer required for function, and downstream DLI optimization.

Enhanced Cell Inactivation and Double-Strand Break Induction in V79 Chinese Hamster Cells by Monochromatic X-Rays at Phosphorus K-Shell Absorption Peak

The cell inactivation and DNA double-strand break (DSB) induction by K-shell ionization of phosphorus atoms and Auger electrons were investigated. Monochromatic X-rays of on and below the phosphorus K-shell absorption peak, 2.153 keV and 2.147 keV were exposed to Chinese hamster lung fibroblast V79 cells. Survival fractions were plotted against exposure, Ψ [nC/kg] and the linear-quadratic model was adapted to estimate the parameters, α and β, of the survival curves. DSB induction rate [DSB/cell/Ψ] was estimated from the measured fractions of induced DNA fragments below 4.6 Mbp (Find(k < 4.6)), which were determined using pulse field gel electrophoresis. As results, cell inactivation and DSB induction rate of on the peak were significantly higher compared to that of the below. However, when converting Ψ to absorbed dose (Gy) of cell nucleus, the enhanced effect was only observed for parameter α, and not for a survival dose (Gy) of 37%, 10%, and 1% nor for a DSB induction rate. Our findings indicate that enhancement of cell inactivation and DSB induction were due to the additional dose delivered to the DNA and more complex DSB lesions were induced due to the release of phosphorus K-shell photoelectrons and Auger electrons.