Low Light Doses Research Articles

Efficiency of Plant UV Irradiation in the Protected Ground

Plants are photosynthetic organisms that depend on sunlight for energy. Besides photosynthetically active radiation (PAR; 400-700 nm), plants are exposed to ultraviolet radiation, which consists of UV-C (below 280 nm), UV-B (280-320 nm) and UV-A (320-400 nm). Plants respond differently to exposure to low or high doses of ultraviolet light, either by stimulating protective mechanisms, or by activating recovery mechanisms to cope with various types of stress. (Research purpose) The research purpose is in providing an overview of the reactions of plants to ultraviolet radiation, make a relationship between the different ranges of ultraviolet radiation, radiation doses and the effect on plants. (Materials and methods) The article presents an analysis of domestic and foreign articles on the effectiveness of plant irradiation with ultraviolet radiation. (Results and discussion) Authors examined the effects of various ranges of ultraviolet radiation on plants. It was found that some properties of plants can be enhanced with small doses of UV-C, but UV-C strongly inhibits plant growth. Exposure to UV-B causes morphological and anatomical changes in plants: an increase in branching, a decrease in internodes, twisting of leaves, a decrease in leaf area, an accumulation of screening pigments, thickening of leaves, redistribution of chlorophyll. The effects of UV-B and UV-C also affect the volatiles emitted by aromatic plants. Ultraviolet-A can improve the nutritional qualities of green crops without adversely affecting plant growth. (Conclusions) It was found that ultraviolet radiation, depending on the range, has both a harmful effect on plants and a beneficial one. It was found that with certain doses of ultraviolet radiation, the necessary qualities and properties of plants could be obtained.

Photodynamic inactivation mediated by 5-aminolevulinic acid of bacteria in planktonic and biofilm forms

Bacterial photodynamic inactivation (PDI) employing endogenous production of porphyrins from 5-aminolevulinic acid (ALA) – named ALA-PDI-, is a new promising tool to achieve bacteria control in non-spread infections. The technique combines the action of the porphyrins acting as photosensitisers with light, to produce reactive oxygen species to target the pathogen. To date, some clinical applications of ALA-PDI have been reported although variable responses ranging from total eradication to absence of photokilling were found. ALA-PDI conducted at suboptimal conditions may lead to misleading results and the complexity of haem synthesis in bacteria hinders the optimization of the treatment.The present work aimed to gain insight on the variables affecting ALA-PDI in Gram-positives and Gram-negatives bacteria growing on planktonic and biofilm cultures and to correlate the degree of the response with the amount and type of porphyrin synthesised.Staphylococcus epidermidis and Escherichia coli clinical isolates and Pseudomonas aeruginosa ATCC27853 and Staphylococcus aureus ATCC25923 strains were utilised, and the optimal conditions of concentration and time exposure of ALA, and light dose were set.In both Gram-positive species analysed, a peak of porphyrin synthesis was observed at 1–2 mM ALA in biofilm and planktonic cultures, which fairly correlated with the decrease in the number of CFU after PDI (5 to 7 logs) and porphyrin content was in the same order of magnitude. In addition, ALA-PDI was similarly effective for planktonic and biofilm S. aureus cultures, and more effective in S. epidermidis planktonic cultures at low light doses. Beyond a certain light dose, it was not possible to achieve further photosensitization. Similarly, a plateau of cell death was attained at a certain ALA incubation time. Accumulation of hydrophilic porphyrins at longer incubation periods was observed.The proportion of porphyrins changed as a function of ALA concentration and incubation time in the Gram-positive bacteria, though we did not find a clear correlation between the porphyrin type and PDI response. As a salient feature was the presence of isococroporphyrin isoforms in both Gram-positive and Gram-negative bacteria.Gram-negative bacteria were quite refractory to the treatment: P. aeruginosa was slightly inactivated (4-logs reduction) at 40 mM ALA, whereas E. coli was not inactivated at all. These species accumulated high ALA quantities and the amount of porphyrins did not correlate with the degree of photoinactivation. Our microscopy studies show that porphyrins are not located in the envelopes of Gram-negative bacteria, reinforcing the hypothesis that endogenous porphyrins fail to attack these structures.

Preclinical Evaluation of Cetuximab and Benzoporphyrin Derivative-Mediated Intraperitoneal Photodynamic Therapy in a Canine Model.

Peritoneal carcinomatosis (PC) can occur as an advanced consequence of multiple primary malignancies. Surgical resection, radiation or systemic interventions alone have proven inadequate for this aggressive cancer presentation, since PC still has a poor survival profile. Photodynamic therapy (PDT), in which photosensitive drugs are exposed to light to generate cytotoxic reactive oxygen species, may be an ideal treatment for PC because of its ability to deliver treatment to a depth appropriate for peritoneal surface tumors. Additionally, epidermal growth factor receptor (EGFR) signaling plays a variety of roles in cancer progression and survival as well as PDT-mediated cytotoxicity, so EGFR inhibitors may be valuable in enhancing the therapeutic index of intraperitoneal PDT. This study examines escalating doses of benzoporphyrin derivative (BPD)-mediated intraperitoneal PDT combined with the EGFR-inhibitor cetuximab in a canine model. In the presence or absence of small bowel resection (SBR) and cetuximab, we observed a tolerable safety and toxicity profile related to the light dose received. Additionally, our findings that BPD levels are higher in the small bowel compared with other anatomical regions, and that the risk of anastomotic failure decreases at lower light doses will help to inform the design of similar PC treatments in humans.

Quantitative Phase Dynamics of Cancer Cell Populations Affected by Blue Light

Increased exposition to blue light may induce many changes in cell behavior and significantly affect the critical characteristics of cells. Here we show that multimodal holographic microscopy (MHM) within advanced image analysis is capable of correctly distinguishing between changes in cell motility, cell dry mass, cell density, and cell death induced by blue light. We focused on the effect of blue light with a wavelength of 485 nm on morphological and dynamical parameters of four cell lines, malignant PC-3, A2780, G361 cell lines, and the benign PNT1A cell line. We used MHM with blue light doses 24 mJ/cm2, 208 mJ/cm2 and two kinds of expositions (500 and 1000 ms) to acquire real-time quantitative phase information about cellular parameters. It has been shown that specific doses of the blue light significantly influence cell motility, cell dry mass and cell density. These changes were often specific for the malignant status of tested cells. Blue light dose 208 mJ/cm2 × 1000 ms affected malignant cell motility but did not change the motility of benign cell line PNT1A. This light dose also significantly decreased proliferation activity in all tested cell lines but was not so deleterious for benign cell line PNT1A as for malignant cells. Light dose 208 mJ/cm2 × 1000 ms oppositely affected cell mass in A2780 and PC-3 cells and induced different types of cell death in A2780 and G361 cell lines. Cells obtained the least damage on lower doses of light with shorter time of exposition.

Unimolecular Photodynamic O2-Economizer To Overcome Hypoxia Resistance in Phototherapeutics.

Tumor hypoxia has proven to be the major bottleneck of photodynamic therapy (PDT) to clinical transformation. Different from traditional O2 delivery approaches, here we describe an innovative binary photodynamic O2-economizer (PDOE) tactic to reverse hypoxia-driven resistance by designing a superoxide radical (O2•-) generator targeting mitochondria respiration, termed SORgenTAM. This PDOE system is able to block intracellular O2 consumption and down-regulate HIF-1α expression, which successfully rescues cancer cells from becoming hypoxic and relieves the intrinsic hypoxia burden of tumors in vivo, thereby sparing sufficient endogenous O2 for the PDT process. Photosensitization mechanism studies demonstrate that SORgenTAM has an ideal intersystem crossing rate and triplet excited state lifetime for generating O2•- through type-I photochemistry, and the generated O2•- can further trigger a biocascade to reduce the PDT's demand for O2 in an O2-recycble manner. Furthermore, SORgenTAM also serves to activate the AMPK metabolism signaling pathway to inhibit cell repair and promote cell death. Consequently, using this two-step O2-economical strategy, under relatively low light dose irradiation, excellent therapeutic responses toward hypoxic tumors are achieved. This study offers a conceptual while practical paradigm for overcoming the pitfalls of phototherapeutics.

PO2-weighted imaging in vivo by delayed fluorescence of intracellular Protoporphyrin IX

In this Letter, we report on the imaging of tissue oxygen by means of time-gated wide-field lifetime imaging of the microsecond-scale delayed fluorescence from Protoporphyrin IX (PpIX). PpIX is produced endogenously from a clinically used δ -aminolevulinic acid (5-ALA) by most cells. The lifetime of delayed fluorescence is lengthened by the absence of oxygen to quench its triplet state. Here, the images of the delayed emission from the human skin and the skin of nude mice were captured after topical application of 5-ALA, using a pulsed 635 nm excitation. Macroscopic lifetime-based images were found to be associated with the tissue oxygenation and obtained with a low light dose ( ∼ 10 m J / c m 2 ) and “capture-times” of a few seconds.

Method for Investigation of Photobiological Effects of Light on Human Skin Cells Mediated by Low Doses of Light.

Driven by evolution, human skin cells have developed an extraordinary ability both to sense and to respond to the photons of sunlight through a plethora of photobiological interactions, activating intracellular signalling cascades and regulating skin cells homeostasis. It has recently been reported that some of these photobiological responses triggered by low levels of light (or the so-called photobiomodulation) could initiatebeneficial therapeutic effects. Identification of these effective light-based therapeutic solutions requires in-depth understanding of the parameter space. The physical, biological, and chemical conditions that need to be fulfilled to facilitate such positive photobiological effects areto be carefully deciphered. Here, we provide the protocols that were specifically developed to investigate multidimensional parameter space driving photobiological interactions triggered by light (photobiomodulation) in the skin cells. The approach is based on the so-called design of experiment (DoE), a statistical method, which allows for the investigation of multidimensional parameters landscapes. This goes hand in hand with sharing practical tips for the design of light-based devices inducing these effects. To exemplify practical applications of the developed methods and light-based devices, we disclose experimental data sets and emphasize robustness and reproducibility of the results.

Ru(ii) photosensitizers competent for hypoxic cancers via green light activation.

A family of five heteroleptic complexes [Ru(C^N)(N^N)2][PF6] (HC^N = methyl 1-butyl-2-arylbenzimidazolecarboxylate; N^N = polypyridine) has been synthesized to act as biologically-compatible green light photosensitizers (PSs) with phototherapeutic indexes (PIs) up to higher than 700 under hypoxia (2% O2) in HeLa cancer cells under short time of irradiation.

Singlet oxygen generation of subphthalocyanine-fused dimer and trimer

Subphthalocyanine (SubPc) macrocycles are known as an interesting class of nonplanar aromatic dyes. Despite documented high fluorescence and singlet oxygen quantum yields, the properties of SubPcs in photodynamic therapy (PDT) are underestimated, because their absorption bands do not reach a significant wavelength range. With this in mind, we combined a SubPc ring and a SubPc ring by introducing a common benzene ring and obtained a SubPc dimer (2) and trimer (3) with the Q-band at the near-IR region, owing to the expansion of the [Formula: see text] electron conjugated system. In this study, we reported1O2generation abilities of 2 and 3based on the applied absolute singlet oxygen quantum yields ([Formula: see text]absolute). Subsequent research revealed that 2 and 3 showed the potential to generate1O2to not only in toluene but also in DMSO. Although the photocytotoxicity of 2 and 3 were investigated upon photo-irradiation with a low light dose of approximately 1.5 J/cm2, 2 and 3 showed almost negligible toxic properties toward HEp2 cells.

Immune Responses after Vascular Photodynamic Therapy with Redaporfin.

Photodynamic therapy (PDT) relies on the administration of a photosensitizer (PS) that is activated, after a certain drug-to-light interval (DLI), by the irradiation of the target tumour with light of a specific wavelength absorbed by the PS. Typically, low light doses are insufficient to eradicate solid tumours and high fluence rates have been described as poorly immunogenic. However, previous work with mice bearing CT26 tumours demonstrated that vascular PDT with redaporfin, using a low light dose delivered at a high fluence rate, not only destroys the primary tumour but also reduces the formation of metastasis, thus suggesting anti-tumour immunity. This work characterizes immune responses triggered by redaporfin-PDT in mice bearing CT26 tumours. Our results demonstrate that vascular-PDT leads to a strong neutrophilia (2–24 h), systemic increase of IL-6 (24 h), increased percentage of CD4+ and CD8+ T cells producing IFN-γ or CD69+ (2–24 h) and increased CD4+/CD8+ T cell ratio (2–24 h). At the tumour bed, T cell tumour infiltration disappeared after PDT but reappeared with a much higher incidence one day later. In addition, it is shown that the therapeutic effect of redaporfin-PDT is highly dependent on neutrophils and CD8+ T cells but not on CD4+ T cells.

Water-Soluble Chlorin/Arylaminoquinazoline Conjugate for Photodynamic and Targeted Therapy.

A new water-soluble conjugate, consisting of a chlorin-e6 photosensitizer part, a 4-arylaminoquinazoline moiety with affinity to epidermal growth factor receptors, and a hydrophilic β-d-maltose fragment, was synthesized starting from methylpheophorbide-a in seven steps. The prepared conjugate exhibited low levels of dark cytotoxicity and pronounced photoinduced cytotoxicity at submicromolar concentrations in vitro, with an IC50(dark)/IC50(light) ratio of ∼368 and a singlet oxygen quantum yield of about 20%. In tumor-bearing Balb/c nude mice, conjugate 1 preferentially accumulates in the tumor tissue. Irradiation of the nude mice bearing A431 xenograft tumors after intravenous administration of the prepared conjugate with a relatively low light dose (50 J/cm2) produced an excellent therapeutic effect with profound tumor regression and low systemic toxicity.

Membrane-Anchoring Photosensitizer with Aggregation-Induced Emission Characteristics for Combating Multidrug-Resistant Bacteria.

Traditional photosensitizers (PSs) show reduced singlet oxygen (1 O2 ) production and quenched fluorescence upon aggregation in aqueous media, which greatly affect their efficiency in photodynamic therapy (PDT). Meanwhile, non-targeting PSs generally yield low efficiency in antibacterial performance due to their short lifetimes and small effective working radii. Herein, a water-dispersible membrane anchor (TBD-anchor) PS with aggregation-induced emission is designed and synthesized to generate 1 O2 on the bacterial membrane. TBD-anchor showed efficient antibacterial performance towards both Gram-negative (Escherichia coli) and Gram-positive bacteria (Staphylococcus aureus). Over 99.8 % killing efficiency was obtained for methicillin-resistant S. aureus (MRSA) when they were exposed to 0.8 μm of TBD-anchor at a low white light dose (25 mW cm-2 ) for 10 minutes. TBD-anchor thus shows great promise as an effective antimicrobial agent to combat the menace of multidrug-resistant bacteria.

Membrane‐Anchoring Photosensitizer with Aggregation‐Induced Emission Characteristics for Combating Multidrug‐Resistant Bacteria

AbstractTraditional photosensitizers (PSs) show reduced singlet oxygen (1O2) production and quenched fluorescence upon aggregation in aqueous media, which greatly affect their efficiency in photodynamic therapy (PDT). Meanwhile, non‐targeting PSs generally yield low efficiency in antibacterial performance due to their short lifetimes and small effective working radii. Herein, a water‐dispersible membrane anchor (TBD‐anchor) PS with aggregation‐induced emission is designed and synthesized to generate 1O2 on the bacterial membrane. TBD‐anchor showed efficient antibacterial performance towards both Gram‐negative (Escherichia coli) and Gram‐positive bacteria (Staphylococcus aureus). Over 99.8 % killing efficiency was obtained for methicillin‐resistant S. aureus (MRSA) when they were exposed to 0.8 μm of TBD‐anchor at a low white light dose (25 mW cm−2) for 10 minutes. TBD‐anchor thus shows great promise as an effective antimicrobial agent to combat the menace of multidrug‐resistant bacteria.

An iridium(III)-palladium(II) metal-organic cage for efficient mitochondria-targeted photodynamic therapy

An Ir8Pd4-heteronuclear metal-organic cage (MOC-51) was assembled from bipodal metalloligand [Ir(ppy)2(qpy)(BF4)] (qpy = 4,4′:2′,2″:4″,4‴-quaterpyridine; ppy = 2-phenylpridine) with Pd(II) salt. The cubic barrel shaped MOC shows one-photon and two-photon excited deep-red emission, as well as large singlet oxygen quantum yields under visible light irradiation, therefore exhibiting great potentials in organelles-targeted cell imaging and photodynamic therapy (PDT). Compared with the Ir(III) metalloligand, the Ir8Pd4-MOC showed less dark toxicity and higher mitochondria-targeting efficiency. The localization in mitochondria overcomes the limitation of short lifetime and diffusion distance of ROS in cell, thus improved PDT effect can be obtained in low light dose usage of the MOC. This study presents the first case of Ir-based metal-organic cages for bio-applications in successful integration of imaging diagnosis and photodynamic therapy

Pulsed Laser Activation of Carbene Bioadhesive Boosts Bonding Strength

AbstractLight‐activated tissue adhesives are limited to low light doses (50 J) and intensities (<1 W cm−2) due to photo‐to‐thermal heat generation. Low intensities have the disadvantage of limited penetration depths with retarded crosslinking kinetics, which impairs carbene‐based crosslinking strategies that compete with nitrogen evolution and gas nucleation. These limitations are circumvented by a trade‐off between high‐intensity activation while reducing the exposure surface area. Continuous or pulsed activation by line scanning the carbene precursor adhesive allows curing a higher surface area/volume ratio while preventing localized heat generation. By optimizing irradiation with a pulsed laser scan, the adhesion strength is improved by 17‐fold over ultraviolet A (UVA) light emitting diodes (LEDs) and is on par with bioadhesive gold standard of topical cyanoacrylates. Overall, this improved method of photo‐activation applies to other industrial and clinical photocuring adhesives where limits on UVA dose constrain exposure intensities.

Benzo[a]phenoselenazine-based NIR photosensitizer for tumor-targeting photodynamic therapy via lysosomal-disruption pathway

The ultimate goal of cancer therapy is to develop antitumor agents that will destroy selectively tumor cells while sparing the health cell of the patient. Herein, we reported a novel tumor-specific and lysosome dual-targeted NIR photosensitizer, Se-Biotin, by conjugating a biotin ligand into benzo[a]phenoselenazinium derivative dye for selective destruction of tumor cells. Attractively, co-culture model showed that Se-Biotin could selectively target to and retain in biotin receptor-overexpressed tumor cells, which as a result significantly minimized the side effects toward normal cells. As confirmed by the in vitro anticancer mechanism, after cellular internalization, upon irradiation, the effective1O2 generation (ΦΔ = 0.69)severely disrupted the lysosomalintegrity and subsequently led to apoptotic cell death. Benefiting from these merits, Se-Biotin successfully achieved a superior anticancer performance with the IC50 as low as 85 nM, only under a low light dose irradiation (12 J/cm2, 660 nm). Therefore, these result demonstrated that Se-Biotin will be a promising PDT agent for targeted cancer photodynamic therapy.

Increasing Resolution in Live Cell Microscopy by Structured Illumination (SIM)

In the context of various approaches to super-resolution microscopy, structured illumination microscopy (SIM) offers several advantages: it needs rather low light doses (with a low risk of phototoxicity or photobleaching), is comparably fast and flexible concerning the use of microscopes, objective lenses and cameras, and has potential for 3D imaging. This paper describes an experimental setup for SIM with first diffraction orders of a spectral light modulator (SLM) creating an interference pattern in two dimensions. We kept this system rather compact with a comparably large illuminated object field, validated it with nano-beads and applied it further to living cells for imaging the cytoskeleton, mitochondria or cell nuclei with a resolution slightly above 100 nm. Its advantages, challenges and limitations—concerning cameras, acquisition time, depth of imaging, light exposure, and combining it with further super-resolving methods—are discussed.

Sunlight-Driven Wearable and Robust Antibacterial Coatings with Water-Soluble Cellulose-Based Photosensitizers.

Herein, a simple, effective, and general strategy is demonstrated to obtain a water-soluble and nontoxic cellulose-based photosensitizer (CPS) with enhanced photodynamic antibacterial activity through introducing protoporphyrin IX (PpIX) and quaternary ammonium salt (QAS) groups onto the cellulose backbone. The synergistic effect of the anchoring and diluting effect of the cellulose backbone and the electrostatic repulsion between QAS groups effectively inhibit the π-π stacking of PpIX groups, thus the as-prepared CPS exhibits markedly enhanced reactive oxygen species (ROS) yield. Meanwhile, the positively charged QAS groups endow the CPS with water-solubility and a strong attractive force to bacteria. As a result, the CPS can rapidly and efficiently kill drug-resistant bacteria strains, including E. coli and S. aureus, with a low light dose (2.4 J cm-2 ) and low concentration of PpIX groups (0.35 × 10-6 m). Benefiting from the excellent processability and formability, the CPS is readily applied as a sunlight-driven wearable and robust antibacterial coating by a spray coating and later crosslinking procedure.

A Combination of Visudyne and a Lipid-anchored Liposomal Formulation of Benzoporphyrin Derivative Enhances Photodynamic Therapy Efficacy in a 3D Model for Ovarian Cancer.

A major objective in developing new treatment approaches for lethal tumors is to reduce toxicity to normal tissues while maintaining therapeutic efficacy. Photodynamic therapy (PDT) provides a mechanistically distinct approach to treat tumors without the systemic toxicity of chemotherapy drugs. PDT involves the light-based activation of a small molecule, a photosensitizer (PS), to generate reactive molecular species (RMS) that are toxic to target tissue. Depending on the PS localization, various cellular and subcellular components can be targeted, causing selective photodamage. It has been shown that targeted lysosomal photodamage followed by, or simultaneous with, mitochondrial photodamage using two different PS results in a considerable enhancement in PDT efficacy. Here, two liposomal formulations of benzoporphyrin derivative (BPD): (1) Visudyne (clinically approved) and (2) an in-house formulation entrapping a lipid conjugate of BPD are used in combination with direct PS localization to mitochondria, endoplasmic reticulum and lysosomes, enabling simultaneous photodamage to all three organelles using a single wavelength of light. Building on findings by our group, and others, this study demonstrates, for the first time in a 3D model for ovarian cancer, that BPD-mediated photodestruction of lysosomes and mitochondria/ER significantly enhances PDT efficacy at lower light doses than treatment with either PS formulation alone.

Engineering biocompatible benzodithiophene-based polymer dots with tunable absorptions as high-efficiency theranostic agents for multiscale photoacoustic imaging-guided photothermal therapy.

To date, photoacoustic imaging (PAI) and PAI-guided photothermal therapy (PTT) have been performed for noninvasive cancer diagnosis and precise ablation of tumors. To conduct concurrent PAI and PTT, it is essential to develop theranostic agents with strong optical absorption and high photothermal transfer efficiency. In this study, we have engineered theranostic agents with tunable absorptions based on conjugated polymer dots (Pdots) with different structures via the simple precipitation method. The as-synthesized Pdots exhibit strong absorption, high biocompatibility, and superior stability. In addition, the Pdots demonstrate that they can serve as contrast agents for multiscale PAI in vitro and in vivo. More importantly, a high photothermal conversion efficiency up to 40% is reached under irradiation with LED light, resulting in effective cancer treatment with extremely low light dose. Consequently, they show the potential as imaging-guided therapeutic agents for clinical cancer treatment and various biomedical applications.