New insights in photodynamic inactivation of Leishmania amazonensis: A focus on lipidomics and resistance.

The emergence of drug resistance in cutaneous leishmaniasis (CL) has become a major problem over the past decades. The spread of resistant phenotypes has been attributed to the wide misuse of current antileishmanial chemotherapy, which is a serious threat to global health. Photodynamic therapy (PDT) has been shown to be effective against a wide spectrum of drug-resistant pathogens. Due to its multi-target approach and immediate effects, it may be an attractive strategy for treatment of drug-resistant Leishmania species. In this study, we sought to evaluate the activity of PDT in vitro using the photosensitizer 1,9-dimethyl methylene blue (DMMB), against promastigotes of two Leishmania amazonensis strains: the wild-type (WT) and a lab induced miltefosine-resistant (MFR) strain. The underlying mechanisms of DMMB-PDT action upon the parasites was focused on the changes in the lipid metabolism of both strains, which was conducted by a quantitative lipidomics analysis. We also assessed the production of ROS, mitochondrial labeling and lipid droplets accumulation after DMMB-PDT. Our results show that DMMB-PDT produced high levels of ROS, promoting mitochondrial membrane depolarization due to the loss of membrane potential. In addition, both untreated strains revealed some differences in the lipid content, in which MFR parasites showed increased levels of phosphatidylcholine, hence suggesting this could also be related to their mechanism of resistance to miltefosine. Moreover, the oxidative stress and consequent lipid peroxidation led to significant phospholipid alterations, thereby resulting in cellular dysfunction and parasite death. Thus, our results demonstrated that DMMB-mediated PDT is effective to kill L. amazonensis MFR strain and should be further studied as a potential strategy to overcome antileishmanial drug resistance.

On-Chip Optical Trapping with High NA Metasurfaces.

Optical trapping of small particles typically requires the use of high NA microscope objectives. Photonic metasurfaces are an attractive alternative to create strongly focused beams for optical trapping applications in an integrated platform. Here, we report on the design, fabrication, and characterization of optical metasurfaces with a numerical aperture up to 1.2 and trapping stiffness greater than 400 pN/µm/W. We demonstrate that these metasurfaces perform as well as microscope objectives with the same numerical aperture. We systematically analyze the impact of the metasurface dimension on the trapping performance and show efficient trapping with metasurfaces with an area as small as 0.001 mm2. Finally, we demonstrate the versatility of the platform by designing metasurfaces able to create multisite optical tweezers for the trapping of extended objects.

Evaluation of dual application of photodynamic therapy-PDT in Candida albicans.

This study aimed to evaluate, in vitro, the efficacy of photodynamic therapy - PDT using dimethyl methylene blue zinc chloride double salt (DMMB) and red LED light on planktonic cultures of Candida albicans. The tests were performed using the ATCC 90,028 strain grown at 37 °C for 24 h, according to a growth curve of C. albicans. The colonies were resuspended in sterile saline adjusted to a concentration of 2 × 108 cells / mL, with three experimental protocols being tested (Protocol 1, 2 and 3) with a fixed concentration of 750 ɳg/mL obtained through the IC50, and energy density 20 J/cm2. Protocol 1 was carried out using conventional PDT, Protocol 2 was applied double PDT in a single session, and Protocol 3 was applied double PDT in two sessions with a 24 h interval. The results showed logarithmic reductions of 3 (4.252575 ± 0.068526) and 4 logs (2.669533 ± 0.058592) of total fungal load in protocols 3 and 2 respectively in comparison to the Control (6.633547 ± 0.065384). Our results indicated that double application in a single session of PDT was the most effective approach for inhibiting the proliferation of Candida albicans (99.991% inhibition).

Two-tier manipulation of holographic information.

Here we demonstrate the two-tier manipulation of holographic information using frequency-selective metasurfaces. Our results show that these devices can diffract light efficiently at designed frequency and environmental conditions. By changing the frequency and refractive index of the surrounding environment, the metasurfaces produce two different holographic images. We anticipate that these environmental dependent, frequency-selective metasurfaces will have practical applications in holographic encryption and sensing.

Production and viscosity of Xanthan Gum are increased by LED irradiation of X. campestris cultivated in medium containing produced water of the oil industry.

Oil recovery is a challenge and microbial enhanced oil recovery is an option. We theorized that the use of produced water (PW) with photo-stimulation could influence both production and viscosity of Xanthan gum. This study aimed at the evaluation of the effect of photo-stimulation by λ630 ± 1 ηm LED light on the biosynthesis of Xanthan gum produced by Xanthomonas campestris IBSBF 2103 strain reusing PW of the oil industry. We assessed the effect of photo-stimulation by LED light (λ630 nm) on the biosynthesis of Xanthan gum produced by X. campestris in medium containing produced water. Different energy densities applied during the microbial growth phase were tested. The highest production was achieved when using 12 J/cm2 LED light (p < 0.01). Three protocols were assessed: Non-irradiated (Control), Irradiation with LED light during the growth phase (LEDgrowth) and Irradiation with LED light during both growth and production phases (LED growth+production). Both the amount and viscosity of the xanthan gum was significantly higher (p < 0.01) in the group LEDgrowth+production. The study showed that LED irradiation (λ630 ± 1 ηm) during both the growth and production phases of the biopolymer increased both the production and viscosity of Xanthan gum.

Organic light emitting diode for in vitro antimicrobial photodynamic therapy of Candida strains.

Organic light emitting diodes (OLEDs) are very attractive light sources because they are large area emitters, and can in principle be deposited on flexible substrates. These features make them suitable for ambulatory photodynamic therapy (PDT). A few reports of in vitro or in vivo OLED based PDT studies for cancer or microbial inhibition are published but to our best knowledge, none against yeasts. Yeast infections are a significant health risk, especially in low income countries with limited medical facilities. In this work, OLED-based antimicrobial PDT (aPDT), using methylene blue (MB) as photosensitizer (PS), is studied to inactivate opportunistic yeast of four Candida strains of two species: Candida albicans and Candida tropicalis. Before aPDT experiments, fluconazole-resistance was evaluated for all strains, showing that both strains of C. tropicalis were resistant and both strains of C. albicans were sensitive to it. We found that 3 repetitive irradiations work better than a single dose while keeping the total fluence constant, and that this result applies whether or not the strains are resistant to fluconazole.

False spectra formation in the differential two-channel scheme of the laser Doppler flowmeter.

Noise in the differential two-channel scheme of a classic laser Doppler flowmetry (LDF) instrument was studied. Formation of false spectral components in the output signal due to beating of electrical signals in the differential amplifier was found out. The improved block-diagram of the flowmeter was developed allowing to reduce the noise.

BV-2 microglial cells sense micro-nanotextured silicon surface topology.

Artificial biomimetic substrates provide useful models for studying cell adhesion, signaling, and differentiation. This article describes biological interactions with a new type of tunable, micro-nanotextured silicon substrate, generated by irradiation of a hydrogenated amorphous silicon film with a large beam, excimer laser (248 nm). In this study, we demonstrate that BV-2 microglial cells can sense differences in laser processed silicon surface topology over the range of 30 nm to 2 µm, where they undergo marked morphogenic changes with increasing feature size. The cells adopt a more elongated shape in the presence of the modified surface structure and exhibit increased levels of actin-rich microdomains, suggesting enhanced adhesion. The excimer laser modification of hydrogenated amorphous silicon to generate micro-nanostructures realizes large area benefits as well as providing a biomaterial where the external and internal structure can be altered and tuned for various applications.