Genetic Transformation of Plasmid DNA into Escherichia coli Using High Frequency Electromagnetic Energy.

We present a novel technique of genetic transformation of bacterial cells mediated by high frequency electromagnetic energy (HF EME). Plasmid DNA, pGLO (5.4 kb), was successfully transformed into Escherichia coli JM109 cells after exposure to 18 GHz irradiation at a power density between 5.6 and 30 kW m-2 for 180 s at temperatures ranging from 30 to 40 °C. Transformed bacteria were identified by the expression of green fluorescent protein (GFP) using confocal scanning microscopy (CLSM) and flow cytometry (FC). Approximately 90.7% of HF EME treated viable E. coli cells exhibited uptake of the pGLO plasmid. The interaction of plasmid DNA with bacteria leading to transformation was confirmed by using cryogenic transmission electron microscopy (cryo-TEM). HF EME-induced plasmid DNA transformation was shown to be unique, highly efficient, and cost-effective. HF EME-induced genetic transformation is performed under physiologically friendly conditions in contrast to existing techniques that generate higher temperatures, leading to altered cellular integrity. This technique allows safe delivery of genetic material into bacterial cells, thus providing excellent prospects for applications in microbiome therapeutics and synthetic biology.

Translocation and fate of nanospheres in pheochromocytoma cells following exposure to synchrotron-sourced terahertz radiation.

The routes by which foreign objects enter cells is well studied; however, their fate following uptake has not been explored extensively. Following exposure to synchrotron-sourced (SS) terahertz (THz) radiation, reversible membrane permeability has been demonstrated in eukaryotic cells by the uptake of nanospheres; nonetheless, cellular localization of the nanospheres remained unclear. This study utilized silica core-shell gold nanospheres (AuSi NS) of diameter 50 ± 5 nm to investigate the fate of nanospheres inside pheochromocytoma (PC 12) cells following SS THz exposure. Fluorescence microscopy was used to confirm nanosphere internalization following 10 min of SS THz exposure in the range 0.5-20 THz. Transmission electron microscopy followed by scanning transmission electron microscopy energy-dispersive spectroscopic (STEM-EDS) analysis was used to confirm the presence of AuSi NS in the cytoplasm or membrane, as single NS or in clusters (22% and 52%, respectively), with the remainder (26%) sequestered in vacuoles. Cellular uptake of NS in response to SS THz radiation could have suitable applications in a vast number of biomedical applications, regenerative medicine, vaccines, cancer therapy, gene and drug delivery.

The Effects of mmW and THz Radiation on Dry Eyes: A Finite-Difference Time-Domain (FDTD) Computational Simulation Using XFdtd.

The importance of investigating the health effects of RF radiation on the cornea cannot be overstated. This study aimed to address this need by utilizing a mathematical simulation to examine the absorption of millimeter wave (mmW) and terahertz (THz) waves by the cornea, considering both normal and pathological conditions. The simulation incorporated variations in tear film thickness and hydration levels, as these factors play a crucial role in corneal health. To assess the impact of RF radiation on the cornea, the study calculated temperature rises, which indicate heating effects for both dry and normal eyes. XFdtd, a widely used commercial software based on the Finite-Difference Time Domain (FDTD) method, was employed to evaluate the radiation absorption and resulting temperature changes. The outcomes of this study demonstrated a crucial finding, i.e., that changes in the water ratio and thickness of the tear film, which are associated with an increased risk of dry eye syndrome, directly impact the absorption of mmW and THz waves by the cornea. This insight provides valuable evidence supporting the interconnection between tear film properties and the vulnerability of the cornea to RF radiation.

The Impact of Base Cell Size Setup on the Finite Difference Time Domain Computational Simulation of Human Cornea Exposed to Millimeter Wave Radiation at Frequencies above 30 GHz.

Mobile communication has achieved enormous technology innovations over many generations of progression. New cellular technology, including 5G cellular systems, is being deployed and making use of higher frequencies, including the Millimetre Wave (MMW) range (30-300 GHz) of the electromagnetic spectrum. Numerical computational techniques such as the Finite Difference Time Domain (FDTD) method have been used extensively as an effective approach for assessing electromagnetic fields' biological impacts. This study demonstrates the variation of the accuracy of the FDTD computational simulation system when different meshing sizes are used, by using the interaction of the critically sensitive human cornea with EM in the 30 to 100 GHz range. Different approaches of base cell size specifications were compared. The accuracy of the computation is determined by applying planar sensors showing the detail of electric field distribution as well as the absolute values of electric field collected by point sensors. It was found that manually defining the base cell sizes reduces the model size as well as the computation time. However, the accuracy of the computation decreases in an unpredictable way. The results indicated that using a cloud computing capacity plays a crucial role in minimizing the computation time.

Investigating the Impact of Synchrotron THz Radiation on the Corneal Hydration Using Synchrotron THz-Far Infrared Beamline.

Due to increasing interest in imaging, industrial, and the development of wireless communication operating at THz frequencies, it is crucial to ascertain possible health impacts arising from exposure to THz radiation. This paper reports on the pilot study of transmittance and absorbance spectra of the porcine cornea following THz frequency irradiation at a synchrotron THz/Far-IR beamline. The exposure period was 4 hours. One cornea was exposed to the radiation, with a second cornea acting as a control. An Attenuated Total Reflection (ATR) apparatus was used, and the frequency range of 2.4 to 8 THz was selected to evaluate any changes. It was found that the synchrotron THz radiation intensities are too low to produce induced corneal injury, but may lead to subtle changes in the state of water. Our results suggest that THz spectroscopy is a promising modality for corneal tissue hydration sensing.

Tissue Characterization Using Synchrotron Radiation at 0.7 THz to 10.0 THz with Extended ATR Apparatus Techniques.

The attenuated total reflection (ATR) apparatus, with an added partial reflection/partial transmission mode, was used to demonstrate a novel way of characterizing water-based substances at 0.7 to 10.0 THz at the Australian Synchrotron THz-far infrared beamline. The technique utilized a diamond-crystal-equipped ATR to track temperature-dependent changes in reflectance. A "crossover flare" feature in the spectral scan was noted, which appeared to be a characteristic of water and water-dominated compounds. A "quiet zone" feature was also seen, where no temperature-dependent variation in reflectance exists. The variation in these spectral features can be used as a signature for the presence of bound and bulk water. The method can also potentially identify the presence of fats and oils in a biological specimen. The technique requires minimal sample preparation and is non-destructive. The presented method has the promise to provide a novel, real-time, low-preparation, analytical method for investigating biological material, which offers avenues for rapid medical diagnosis and industrial analysis.

Computational phantom study of frozen melanoma imaging at 0.45 terahertz.

Terahertz radiation (THz) is highly absorbed by liquid water. This creates the possibility of medical imaging on the basis of the water content difference between normal and diseased tissue. The effective penetration of THz is limited, however, to a tissue depth of 0.2-0.3 mm at body temperature. A unique feature of the 0.1-2.0 THz frequency is that there is a high disparity between liquid water absorption and ice absorption, with ice being 100 times more permeable to the radiation than liquid water. This results in 90% of the radiation surviving to 1.0 mm in ice, permitting the imaging of frozen tissues to a depth of 5.0 mm. This method is practical as an in vivo procedure before or during surgical excision. Finite difference time domain (FDTD) computational modeling of frozen normal skin and frozen melanoma was undertaken using tissue phantoms. The study suggests that sufficient contrast exists to differentiate normal frozen skin and melanoma on the basis of the difference of water content alone. When the melanin pigment in melanomas is modeled as a significant absorber of THz, the contrast changes. Based on the modeling, further exploration of the "THz-skin freeze" imaging technique is justified. In the modeling, the boundary between the frozen tissue and non-frozen tissue is shown to be strongly reflective. If the reflective properties of the boundary are substantiated, the "THz-skin freeze" technique will have applications in other areas of skin diagnostics and therapeutics. Bioelectromagnetics. 40:118-127, 2019. © 2019 Bioelectromagnetics Society.

THz Filters Made by Laser Ablation of Stainless Steel and Kapton Film.

THz band-pass filters were fabricated by femtosecond-laser ablation of 25-µm-thick micro-foils of stainless steel and Kapton film, which were subsequently metal coated with a ∼70 nm film, closely matching the skin depth at the used THz spectral window. Their spectral performance was tested in transmission and reflection modes at the Australian Synchrotron's THz beamline. A 25-µm-thick Kapton film performed as a Fabry-Pérot etalon with a free spectral range (FSR) of 119 cm-1, high finesse Fc≈17, and was tuneable over ∼10µm (at ∼5 THz band) with ß=30∘ tilt. The structure of the THz beam focal region as extracted by the first mirror (slit) showed a complex dependence of polarisation, wavelength and position across the beam. This is important for polarisation-sensitive measurements (in both transmission and reflection) and requires normalisation at each orientation of linear polarisation.

Spectroscopy of excised skin patches exposed to THz and far-IR radiation.

Applications of far infrared (Far-IR) and terahertz (THz) radiation in areas such as healthcare and security are fast-growing. As a consequence, humans and the environment are becoming more exposed to mm-wave and Far-IR radiation than previously. We examined typical skin-care and sunscreen ingredients and propitiatory products with transmission FTIR, ATR-FTIR and THz-time domain spectroscopy (THz-TDS) methods using fresh and dehydrated toad and fresh human skin samples for their absorption properties in these frequency ranges. The skin hydration compounds glycerol and sorbitol have comparable absorption characteristics to physiological bulk water. Products containing these and similar hydrating compounds have significant Far-IR absorption characteristics. The sunscreen ingredients ZnO (20 micron), TiO2 (mesh 325), and graphene platelet demonstrate a generally poor Far-IR absorbance, with TiO2 displaying some frequency-specific absorption in the 3-6 THz and 12 THz regions. The Far-IR absorbance of proprietary sunscreens was, however, shown not to be significant. The absorption properties of melanin, collagen, bound water, and other constituents are significant in dehydrated skin samples but are not of the same order of importance as the hydrating agents examined.

Translocation of silica nanospheres through giant unilamellar vesicles (GUVs) induced by a high frequency electromagnetic field.

Membrane model systems capable of mimicking live cell membranes were used for the first time in studying the effects arising from electromagnetic fields (EMFs) of 18 GHz where membrane permeability was observed following exposure. A present lack of understanding of the mechanisms that drive such a rapid change in membrane permeabilization as well as any structural or dynamic changes imparted on biomolecules affected by high-frequency electromagnetic irradiation limits the use of 18 GHz EMFs in biomedical applications. A phospholipid, 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) labelled with a fluorescent marker 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N-(lissamine rhodamine B sulfonyl) (rhodamine-DOPE) was used in constructing the giant unilamellar vesicles (GUVs). After three cycles of exposure, enhanced membrane permeability was observed by the internalisation of hydrophilic silica nanospheres of 23.5 nm and their clusters. All-atom molecular dynamics simulations of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) membranes exposed to high frequency electric fields of different field strengths showed that within the simulation timeframe only extremely high strength fields were able to cause an increase in the interfacial water dynamics characterized by water dipole realignments. However, a lower strength, high frequency EMF induced changes of the water hydrogen bond network, which may contribute to the mechanisms that facilitate membrane permeabilization in a longer timeframe.

Computer simulation study of the penetration of pulsed 30, 60 and 90 GHz radiation into the human ear.

There is increasing interest in applications which use the 30 to 90 GHz frequency range, including automotive radar, 5 G cellular networks and wireless local area links. This study investigated pulsed 30-90 GHz radiation penetration into the human ear canal and tympanic membrane using computational phantoms. Modelling involved 100 ps and 20 ps pulsed excitation at three angles: direct (orthogonal), 30° anterior, and 45° superior to the ear canal. The incident power flux density (PD) estimation was normalised to the International Commission on Non-Ionizing Radiation Protection (1998) standard for general population exposure of 10 Wm-2 and occupational exposure of 50 Wm-2. The PD, specific absorption rate (SAR) and temperature rise within the tympanic membrane was highly dependent on the incident angle of the radiation and frequency. Using a 30 GHz pulse directed orthogonally into the ear canal, the PD in the tympanic membrane was 0.2% of the original maximal signal intensity. The corresponding PD at 90 GHz was 13.8%. A temperature rise of 0.032° C (+20%, -50%) was noted within the tympanic membrane using the equivalent of an occupational standard exposure at 90 GHz. The central area of the tympanic membrane is exposed in a preferential way and local effects on small regions cannot be excluded. The authors strongly advocate further research into the effects of radiation above 60 GHz on the structures of the ear to assist the process of setting standards.

Computational absorption and reflection studies of normal human skin at 0.45 THz.

Applications using terahertz (THz) frequency radiation will inevitably lead to increased human exposure. The power density and specific absorption rate (SAR) simulations of thin skin at 0.45 THz show the bulk of the energy being absorbed in the upper stratum spinosum, and the maximal temperature rise is in the lower stratum spinosum. There are regions of SAR increase of 100% above the local average at the stratum spinosum/stratum basale boundary. The dead Stratum Corneum layer protects underlying tissues in thick skin. Reflection studies suggest that acute angles and the use of polarised incident radiation may enhance the assessment of diabetic neuropathy.

Computational simulations of the penetration of 0.30 THz radiation into the human ear.

There is an increasing interest in industrial and security applications and the establishment of wireless communication operating at frequencies of up to 0.30 THz. Soft tissue has a high coefficient of absorption at 0.30 THz and this limits effective penetration of the energy to a depth of 0.2 to 0.4 mm. The capacity of 0.30 THz radiation to access the deeper parts of the ear by diffusing through the ear canal and exposing the tympanic membrane (ear drum) to the radiation has not been studied. Simulations show that, with excitation parallel to the ear canal, the average power flux density within the central tympanic membrane region is 97% of the incident excitation. The structures of the outer ear are highly protective; less than 0.4% of the power flux density is directed at 45° from the parallel reached the same region. Given the sensitivity of the tympanic membrane to mechanical change, in-vivo assessment of the penetration of 0.3 THz into the ear canal is warranted to assess the suitability of the present radiation safety limits and to inform 0.3 THz emitting device deign.