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Establishment of a highly efficient cell factory is imperative for 5-aminolevulinic acid (5-ALA) biomanufacturing. A streamlined workflow is described that enables highly efficient 5-ALA synthase mining. Genome-scale model-guided identification and combination of multiplex targets are reported. An artificial homeostasis was designed for dynamically responding to, and fine-tuning, redox status. Final collaborative optimization resulted in the highest 5-ALA biomanufacturing performance achieved to date. The versatile applications of 5-aminolevulinic acid (5-ALA) across the fields of agriculture, livestock, and medicine necessitate a cost-efficient biomanufacturing process. In this study, we achieved the economic viability of biomanufacturing this compound through a systematic engineering framework. First, we obtained a 5-ALA synthase (ALAS) with superior performance by exploring its natural diversity with divergent evolution. Subsequently, using a genome-scale model, we identified and modified four key targets from distinct pathways in Escherichia coli , resulting in a final enhancement of 5-ALA titers up to 21.82 g/l in a 5-l bioreactor. Furthermore, recognizing that an imbalance of redox equivalents hindered further titer improvement, we developed a dynamic control system that effectively balances redox status and carbon flux. Ultimately, we collaboratively optimized the artificial redox homeostasis system at the transcription level with other cofactors at the feeding level, demonstrating the highest recorded performance to date with a titer of 63.39 g/l for the biomanufacturing of 5-ALA. Graphical abstract [Display omitted] The economic viability of biomanufacturing 5-aminolevulinic acid (5-ALA) was successfully demonstrated in this study, showcasing excellent production performance. The titer reached a record-breaking 63.39 g/l at 44 h, representing the highest reported value to date, with the productivity of 1.44 g/l/h. Although the yield (0.384 mol/mol glucose) was lower than theoretically expected, the significant value of 5-ALA positions our developed cell factory competitively for efficient industrial-scale biomanufacturing. Therefore, no challenges unique to this compound can be identified for full-scale fermentation, particularly considering that our 5-ALA cell factory was derived from a widely utilized Escherichia coli strain. Currently, two primary approaches are used to produce 5-ALA: chemical synthesis and biomanufacturing. Microbial biosynthesis of 5-ALA presents a more facile, environmentally benign, and low-cost alternative. Given that 5-ALA is a non-protein amino acid, we anticipated that the entire biomanufacturing process would be analogous to the biomanufacturing of canonical amino acids. Therefore, although a separation process was not implemented in this study, we believe that our developed 5-ALA cell factory exhibits excellent parameters and represents a highly cost-efficient biomanufacturing process. Within NASA's Technology Readiness Level (TRL) system, we propose that this 5-ALA cell factory has reached TRL 6, indicating a fully functional prototype suitable for demonstration in real production scenarios. A systematic engineering framework was demonstrated to construct a 5-aminolevulinic acid (5-ALA) cell factory, achieving the economic viability of biomanufacturing this compound with an unprecedented production performance (63.39 g/l). This comprehensive framework encompasses enzyme mining, multitarget engineering, artificial homeostasis design, and collaborative optimization. [ABSTRACT FROM AUTHOR]

Cerenkov radiation induced photodynamic therapy (CR-PDT) can tackle the tissue penetration limitation of traditional PDT. However, co-delivery of radionuclides and photosensitizer may cause continuous phototoxicity in normal tissues during the circulation. 5-aminolevulinic acid (ALA) which can intracellularly transform into photosensitive protoporphyrin IX (PpIX) is a cancer-selective photosensitizer with negligible side effect. However, the hydrophilic nature of ALA and the further conversion of PpIX to photoinactive Heme severely hinder the therapeutic benefits of ALA-based PDT. Herein, we developed an 89Zr-labeled, pH responsive ALA and artemisinin (ART) co-loaded liposome (89Zr-ALA-Liposome-ART) for highly selective cancer therapy. 89Zr can serve as the internal excitation source to self-activate PpIX for CR-PDT, and the photoinactive Heme can activate the chemotherapeutic effect of ART. The 89Zr-ALA-Liposome-ART exhibited excellent tumor inhibition capability in subcutaneous 4T1-tumor-bearing Balb/c mice via CR-PDT and chemotherapy. Combined with anti-PD-L1, the 89Zr-ALA-Liposome-ART elicited strong antitumor immunity to against tumor recurrence. A radiolabeled dual drug-loaded responsive liposome was designed to achieve tumor-selective Cerenkov radiation induced photodynamic therapy and combined chemotherapy. [Display omitted] • 89Zr labeled self-illuminated liposome facilitates PDT for deep tumors. • pH responsive release of prodrug ALA reduces the phototoxicity to normal tissue. • Heme generated from exogenous ALA activates the ART for combined chemotherapy. [ABSTRACT FROM AUTHOR]

Modified 5-aminolevulinic acid photodynamic therapy (M-PDT) and isotretinoin (ISO) are effective treatments for moderate to severe acne vulgaris. To evaluate the efficacy and adverse effects of M-PDT and ISO for moderate to severe acne vulgaris. A multicenter, randomized clinical trial was conducted with participants randomly assigned to the M-PDT group (up to 5 weekly sessions following manual comedone extraction) or the ISO group (oral ISO, 0.5 mg/kg/d for 6 months) and followed up to 6-months after therapy. A total of 152 patients were allocated. The overall effective rates in the M-PDT group were significantly higher than the ISO group at 1 month (67.74% vs 10.26%), whereas the opposite was the case 1 month after treatment (75.81% vs 97.44%). Time to achieve 50% lesion improvement in the M-PDT group was significantly less than the ISO group (1 vs 8 weeks). Overall, 70.67% of the ISO group patients experienced systemic side effects such as hepatotoxicity, whereas side effects were skin-limited in the M-PDT group. Limitations of this study included relatively low numbers of participants and high withdrawal rate. M-PDT offers a more rapid onset of improvement, comparable overall efficacy, good tolerability, and comparable durability of response compared with ISO. [ABSTRACT FROM AUTHOR]

Photodynamic therapy, in which photosensitizers locally generate cytotoxic reactive oxygen species, can treat tumor tissue with minimal effects on surrounding normal tissue, but it can be ineffective because of the anoxic tumor microenvironment. Here we developed a strategy to inactivate the mitochondria of tumor cells in order to ensure adequate local oxygen concentrations for photodynamic therapy. We conjugated the photosensitizer 5-aminolevulinic acid to the lipophilic cation triphenylphosphine, which targets mitochondria. Then we packaged the conjugate into nanoparticles that were based on biocompatible bovine serum albumin and coated with folic acid in order to target the abundant folate receptors on the tumor surface. In studies in cell culture and BALB/c mice bearing MCF-7 xenografts, we found that the nanoparticles helped solubilize the cation-photosensitizer conjugate, prolong its circulation, and enhance its photodynamic antitumor effects. We confirmed the ability of the nanoparticles to target tumor cells and their mitochondria using confocal laser microscopy and in vivo assays of pharmacokinetics, pharmacodynamics, and tissue distribution. Our results not only identify a novel nanoparticle system for treating cancer, but they demonstrate the feasibility of enhancing photodynamic therapy by reducing oxygen consumption within tumors. Schematic diagram of FA-BAT-NPs for targeting the mitochondria of breast cancer cells to restore oxygen supply for photodynamic therapy. BAT, Boc-ALA-TPP; BSA, bovine serum albumin; BAT-NPs, BAT-loaded nanoparticles; FA-BAT-NPs, folic acid modified BAT-NPs; FR, folate receptor. [Display omitted] • Inactivating mitochondria of tumor cells to ensure adequate local ROS for photodynamic therapy. • Targeting nanoparticles to the mitochondria of tumor cells to restore oxygen supply for PDT. • Increasing local levels of oxygen by destroying mitochondria within tumor cells. • Triphenylphosphine as a mitochondrial-targeting molecule was conjugated with photosensitizer. • Converting photosensitizer photodynamically into additional ROS. [ABSTRACT FROM AUTHOR]

Hypertrophic scar is a common problem after skin burns or trauma which brings physical, psychological, and cosmetic problems to patients. Photodynamic therapy with 5-aminolevulinic acid (5-ALA) is a promising therapy for hypertrophic scar. However, clinical applications of 5-ALA are limited because of the low permeability of 5-ALA in the skin stratum corneum and the rapid binding of protoporphyrin IX (PpIX) with iron ions, which lead to insufficient PpIX production in target tissues. Herein, a mixture of 5-ALA and DFO (deferoxamine, a special iron chelator) was applied for the treatment of hypertrophic scar. 5-ALA/DFO could efficiently block the biotransformation of PpIX to heme, thus realizing a significant accumulation of photosensitizer. In addition, injection locally into the lesion was applied, which combined with enhanced photodynamic therapy to destroy hypertrophic scar fibroblasts. In vitro experiments showed that 5-ALA/DFO could increase more ROS generation by increasing the accumulation of PpIX, resulting in the apoptosis of hypertrophic scar fibroblasts. Furthermore, 5-ALA/DFO inhibited the proliferation and migration of hypertrophic scar fibroblasts. In vivo study showed that 5-ALA/DFO could effectively inhibit the formation of proliferative scar. Therefore, 5-ALA/DFO has the potential to enhance the photodynamic therapy of 5-ALA and provides a new treatment strategy for hypertrophic scar. Scheme 1. Schematic illustration of the construction of 5-ALA/DFO for enhanced PDT for hypertrophic scar. [Display omitted] [ABSTRACT FROM AUTHOR]

The acute hepatic porphyrias (AHP) are rare, inborn errors of heme-metabolism and include acute intermittent porphyria, hereditary coproporphyria, variegate porphyria, and porphyria due to severe deficiency of 5-aminolevulinic acid dehydratase. Acute intermittent porphyria is the most common type of AHP, with an estimated prevalence of patients with symptoms of approximately 1 in 100,000. The major clinical presentation involves attacks of severe pain, usually abdominal and generalized, without peritoneal signs or abnormalities on cross-sectional imaging. Acute attacks occur mainly in women in their childbearing years. AHP should be considered in the evaluation of all patients, and especially women aged 15–50 years with recurrent severe abdominal pain not ascribable to common causes. The screening tests of choice include random urine porphobilinogen and δ-aminolevulinic acid corrected to creatinine. All patients with elevations in urinary porphobilinogen and/or δ-aminolevulinic acid should initially be presumed to have AHP. The cornerstones of management include discontinuation of porphyrinogenic drugs and chemicals, administration of oral or intravenous dextrose and intravenous hemin, and use of analgesics and antiemetics. Diagnosis of AHP type can be confirmed after initial treatment by genetic testing for pathogenic variants in HMBS, CPOX, PPOX, and ALAD genes. AHP is also associated with chronic symptoms and long-term risk of systemic arterial hypertension, chronic renal and liver disease, and hepatocellular carcinoma. Patients who have recurrent acute attacks (4 or more per year) should be considered for prophylactic therapy with intravenous hemin or subcutaneous givosiran. Liver transplantation is curative and reserved for patients with intractable symptoms who have failed other treatment options. This expert review was commissioned and approved by the American Gastroenterological Association (AGA) Institute Clinical Practice Updates Committee (CPUC) and the AGA Governing Board to provide timely guidance on a topic of high clinical importance to the AGA membership, and underwent internal peer review by the CPUC and external peer review through standard procedures of Gastroenterology. These Best Practice Advice (BPA) statements were drawn from a review of the published literature and from expert opinion. Because systematic reviews were not performed, these BPA statements do not carry formal ratings of the quality of evidence or strength of the presented considerations. Best Practice Advice Statements Women aged 15–50 years with unexplained, recurrent severe abdominal pain without a clear etiology after an initial workup should be considered for screening for an AHP. Initial diagnosis of AHP should be made by biochemical testing measuring δ-aminolevulinic acid, porphobilinogen, and creatinine on a random urine sample. Genetic testing should be used to confirm the diagnosis of AHP in patients with positive biochemical testing. Acute attacks of AHP that are severe enough to require hospital admission should be treated with intravenous hemin, given daily, preferably into a high-flow central vein. In addition to intravenous hemin, management of acute attacks of AHP should include pain control, antiemetics, management of systemic arterial hypertension, tachycardia, and hyponatremia, and hypomagnesemia, if present. Patients should be counseled to avoid identifiable triggers that may precipitate acute attacks, such as alcohol and porphyrinogenic medications. Prophylactic heme therapy or givosiran, administered in an outpatient setting, should be considered in patients with recurrent attacks (4 or more per year). Liver transplantation for AHP should be limited to patients with intractable symptoms and significantly decreased quality of life who are refractory to pharmacotherapy. Patients with AHP should be monitored annually for liver disease. Patients with AHP, regardless of the severity of symptoms, should undergo surveillance for hepatocellular carcinoma, beginning at age 50 years, with liver ultrasound every 6 months. Patients with AHP on treatment should undergo surveillance for chronic kidney disease annually with serum creatinine and estimated glomerular filtration rate. Patients should be counseled on the chronic and long-term complications of AHP, including neuropathy, chronic kidney disease, hypertension, and hepatocellular carcinoma, and need for long-term monitoring. [ABSTRACT FROM AUTHOR]

[Display omitted] Acne is a chronic inflammatory disease of pilosebaceous unit, which can be aggravated by hyperkeratosis of the pilosebaceous unit, excessive secretion of sebum and the proliferation of Propionibacterium acnes (P. acnes). Traditional drug treatment methods commonly exhibit drawbacks, including bacterial resistance and poor transdermal permeability, resulting in suboptimal efficacy and recurrent infections. Herein, we designed a dual drug-loaded microneedles patch to deliver ALA and silver nanoparticles for acne treatment. The comprehensive treatment of acne is achieved by utilizing the safe and efficient advantages of 5-aminolevulinic acid (ALA) photodynamic therapy (PDT), coupled with the superior antibacterial properties of silver nanoparticles. To enhance drug permeability, dissolving microneedles (DMNs) are employed for targeted and efficient drug delivery. In vitro and in vivo experiments demonstrated that DMNs loaded with both AgNPs and ALA-PDT (ALA/AgNPs@DMNs) have significant antibacterial activity, along with superior drug loading capacity and penetration. The application of ALA/AgNPs@DMNs in treating acne-affected rats alleviates excessive epidermal keratinization, effectively inhibits bacterial growth, and significantly improves acne symptoms. This suggests that the "dissolving microneedle" formulation holds promise for the treatment and management of acne. [ABSTRACT FROM AUTHOR]

Hydrogen sulfide (H₂S) is considered as plant growth promoter under heavy metal stress, though its specific effects on photosynthesis are rarely explored. This study investigates the protective effects of exogenous H 2 S donor sodium hydrosulfide (NaHS) on chlorophyll metabolism and photosystem II (PSII) function in 24-day-old bean plants exposed to 10 μM chromium (Cr) stress. Sodium hydrosulfide (100 μM) reduced Cr accumulation in both roots and leaves, leading to restored plant growth. Concomitantly, H₂S mitigated Cr-induced oxidative damages by decreasing reactive oxygen species levels and further enhancing antioxidant scavenging activities. This resulted in significant reductions in Cr-elevated leaf pheophytin and chlorophyllide levels by 59% and 67%, respectively. Furthermore, NaHS application increased levels of porphyrin and its precursor, 5-aminolevulinic acid (5-ALA), in Cr-stressed bean. The up-regulation in chlorophyll biosynthesis was associated with enhanced activities of glutamine synthetase and glutamate synthase, essential for glutamate (precursor of 5-ALA) production, as well as nitrate and nitrite reductase, leading to increased nitric oxide generation. Under Cr stress, H₂S significantly improved the electron transport rate, effective quantum yield of PSII, and photochemical quenching by 112%, 53%, and 38%, respectively, while reducing non-photochemical quenching by 50%. Furthermore, H₂S promoted net CO₂ assimilation and photosynthesis at saturating light, respectively, while reducing stomatal conductance and transpiration to maintain water balance. Exogenous H₂S restored respiration, as indicated by increased light saturation and compensation points in Cr-treated plants. Overall, these findings indicate that H₂S regulates photosynthesis in Cr-stressed bean by modulating nitrogen and chlorophyll metabolism, thereby optimizing PSII efficiency and gas exchange. [Display omitted] • Hydrogen sulfide (H 2 S) reduces chlorophyll breakdown in chromium (Cr)-stressed bean. • H 2 S boosts nitrogen metabolism, improving nitric oxide level in bean exposed to Cr. • Under Cr stress, H 2 S promotes synthesis of 5-aminolevulinic acid and porphyrins. • H 2 S improves chlorophyll levels and photosystem II activity in Cr contaminated bean. • H 2 S regulates stomatal conductance, carbon assimilation, and respiration in Cr stress. [ABSTRACT FROM AUTHOR]

• ALA-PDT significantly improved VLS patients' clinical symptoms and signs. • The side effects of ALA-PDT for VLS were transient and tolerable which led to high patients' satisfaction. • ALA-PDT is a safe and effective treatment for women with intractable VLS. Female vulvar lichen sclerosus (VLS) is a chronic inflammatory skin disease of the vulva and its etiology is unknown. The main clinical symptoms are itching, burning and dyspareunia, and there is a lack of effective treatment. Clinical and follow-up data of women with VLS who underwent 5-Aminolevulinic acid photodynamic therapy (ALA-PDT) from January 2023 to December 2023 in the department of obstetrics and gynecology, Qilu Hospital of Shandong University, were retrospectively analyzed. According to the inclusion and exclusion criteria, 36 patients with VLS who received ineffective conventional treatment (intractable VLS) were enrolled. Objective signs and subjective symptoms of vulvar lesions were recorded before treatment and 6 months after the end of treatment according to corresponding scoring criteria. Quality of life was evaluated using the Dermatology Life Quality Index (DLQI). All patients received six sessions of ALA-PDT treatment and follow-up visits. After ALA-PDT treatment, 24 of 36 (66.67 %) patients' itching symptoms completely disappeared, 10 of 36 (27.78 %) patients' itching symptoms were relieved from severe to mild, and only 2 of 36 (5.56 %) patients' symptoms were not significantly relieved. 16 of 36 (44.4 %) patients' itching symptoms completely disappeared, 9 of 36 (25 %) patients' itching symptoms were relieved from severe to mild, and only 2 of 36 (5.56 %) patients still had severe pain. Compared to 22 patients with dyspareunia before treatment, only 9 patients still had dyspareunia with varying degrees of dyspareunia relief after treatment. Clinical signs improved significantly in the patients after ALA-PDT treatment. The total scores of clinical signs were (5.31 ± 1.67 vs 3.67 ± 1.71) before and after treatment. All patients showed improvement in DLQI after treatment. The main side effects of ALA-PDT were pain, erythema and swelling which were transient and tolerable. All patients were "satisfied" or "very satisfied" with the results of the treatment. ALA-PDT is a safe and effective treatment for women with intractable vulva lichen sclerosus. [ABSTRACT FROM AUTHOR]

• Surgery combined with ALA-PDT is a safe and effective method for the treatment of cutaneous tropical candida infection. • Conventional antifungal therapy relies heavily on drugs, but cutaneous fungal infections (e.g., Candida tropicalis) are highly drug-resistant, and surgery combined with ALA-PDT is a promising alternative for the treatment of refractory cutaneous Candida tropicalis infections. Candidiasis is a common fungal infection, with Candida tropicalis infections being relatively rare. These infections develop into skin fungal infections under conditions such as broad-spectrum antibiotic use, diabetes, and immunosuppression. Traditionally, antifungal drugs have been the primary treatment; however, issues like hepatotoxicity and the need for long-term use persist. Consequently, new treatment approaches are essential. Most previous reports have focused on Candida albicans, with fewer addressing Candida tropicalis. This study reports a case where a patient with a skin Candida tropicalis infection was treated with a combination of surgery and ALA photodynamic therapy(635 nm, 120 J/cm2, 177 mW/cm2). After just one session of this combined treatment, the patient achieved significant clinical remission. Therefore, the combination of surgery and ALA-PDT may offer a safe and effective treatment option. [ABSTRACT FROM AUTHOR]

• Oral administration of 5-aminolevulinic acid enhances photodynamic diagnosis but increases the risk of specific adverse events during bladder tumor resection. • Significant associations found between spinal anesthesia and perioperative hypotension; advanced age and higher BMI with nausea/vomiting and male sex and longer operative times with increased photosensitivity. • Patients using renin-angiotensin system inhibitors showed a higher incidence of postoperative liver dysfunction, suggesting the need for careful monitoring. • The study identifies critical risk factors that could guide preoperative planning and patient counseling to mitigate potential adverse effects in vulnerable populations. • This research underscores the importance of individualized patient assessments when using 5-aminolevulinic acid in surgical procedures for bladder cancer. Oral 5-aminolevulinic acid for transurethral resection of bladder tumor reduces bladder cancer recurrence compared with standard white-light transurethral resection of bladder tumor. However, data regarding risks of adverse events with this drug are unclear. The aim of the present study was to identify risk factors associated with oral 5-aminolevulinic acid induced adverse events in photodynamic diagnosis-transurethral resection of bladder tumor. We studied 104 cases of 5-aminolevulinic acid-photodynamic diagnosis-transurethral resection of bladder tumor from October 2021 to April 2023, administering 20 mg/kg 5-aminolevulinic acid orally at least 2 h pre-surgery. Four major adverse events associated with 5-aminolevulinic acid were selected to identify risk factors for their occurrence, including perioperative hypotension, nausea and/or vomiting, photosensitivity, and liver dysfunction. Univariate and multivariate analyses were conducted to identify the risk factors of those adverse events. Perioperative hypotension (11.5 %), nausea and/or vomiting (37.5 %) photosensitivity (31.7 %), and liver dysfunction (51.9 %) were observed. Multivariate analyses revealed that spinal anesthesia was associated with hypotension (p = 0.02), whereas advanced age (p < 0.01) and higher body mass index (p < 0.01) were associated with nausea and/or vomiting. Also, male sex (p < 0.01) and longer operative time (p = 0.01) were associated with photosensitivity, and renin-angiotensin system inhibitors use was associated with postoperative liver dysfunction (p < 0.01). For elderly male obese patients taking renin-angiotensin system inhibitors, particular attention is needed during the perioperative period of photodynamic diagnosis-transurethral resection of bladder tumor under spinal anesthesia due to the higher risk of onset of the adverse events involved in oral administration of 5-aminolevulinic acid. [ABSTRACT FROM AUTHOR]

Photodynamic therapy (PDT) is an effective cancer treatment option, but it suffers from penetration limit of light, making it available only for superficial and endoscopically accessible cancers. Recently, there have been reports that Cerenkov luminescence originated from radioisotopes can be utilized as an excitation source for PDT without external light illumination. Here, cancer-selective agents, i.e. , (1) clinically available 5-aminolevulinic acid (5-ALA), which promotes cancer metabolism-specific accumulation of protoporphyrin IX (PpIX), and (2) 64Cu-DOTA-trastuzumab, which has HER2-expressing cancer selective uptake, are separately applied as a photosensitizer and an in situ radiator, respectively, to potentiate tumor-specific Cerenkov luminescence energy transfer (CLET) from 64Cu to PpIX for high-precision PDT of cancer. It is shown that the combinational administration and tumor colocalization of 5-ALA and 64Cu-DOTA-trastuzumab exert significant in vitro cytotoxicity (cell viability <9%) as well as in vivo antitumor effects (tumor volume ratio of 0.50 on 14 days post-injection) on HER2-expressing breast and gastric cancer models. This study proves that high-precision treatment regimen using dual-targeted CLET-based PDT is feasible for HER2-expressing cancers. Furthermore, the results offer great potential for clinical translation to the dual-targeted CLET-based PDT because the treatment regimen uses components, 5-ALA and 64Cu-DOTA-trastuzumab, which are already in clinical uses. Administration of 5-aminolevulinic acid and 64Cu-DOTA-trastuzumab results in accumulation of protoporphyrin IX and charged particles within HER2-expressing cancer generating 1O 2 via Cerenkov luminescence energy transfer (CLET) causing cancer cell death. [Display omitted] • Protophorphyrin IX (PpIX) can be excited by 64Cu via Cerenkov luminescence energy transfer (CLET). • Injection of 64Cu-DOTA-trastuzumab and 5-ALA leads to tumor-specific accumulation of charged particle and PpIX, respectively. • Co-administration of 64Cu-DOTA-trastuzumab and 5-ALA results in significant inhibition of tumor growth in a mouse model. • The results provide a novel cancer treatment method based on dual-targeted CLET-based photodynamic therapy. [ABSTRACT FROM AUTHOR]

The ORBEYE (ORB), an innovative 3-dimensional digital exoscope, is an equipped system for fluorescence-guided surgery with 5-aminolevulinic acid. Therefore, this study aimed to verify the characteristics of fluorescence-guided surgery with 5-aminolevulinic acid and excitation light source with ORB. The same operative field of glioblastoma was recorded under blue light (BL) excitation using a conventional microscope (MS) and ORB. For in vitro studies, the energy of 405-nm wavelength light in white light and BL modes of each scope was examined in various focal lengths. To examine the degree of photobleaching with BL for each scope, protoporphyrin IX–soaked filter papers were continuously exposed with BL of an MS and ORB, and the video-recorded red fluorescence intensity was analyzed. The color tone of tumor-induced red fluorescence was remarkably different under each scope. Furthermore, nonfluorescent normal structures without red fluorescence were well recognized under ORB. The energy of 405-nm wavelength light in BL was significantly higher in ORB than that in an MS, especially in the short focal length. With continuous BL excitation to filter papers, the relative red fluorescence intensity of filter papers was significantly decreased over time in ORB than in an MS. In low protoporphyrin IX concentration, the difference was more significant. With ORB, the good visibility due to BL energy as compared with an MS might improve the surgical manipulation even in BL mode. However, the weak fluorescent tissue and short focal length should be carefully considered because photobleaching might be critical for FGS. [ABSTRACT FROM AUTHOR]

5-Aminolevulinic acid (ALA), an antioxidant existing in plants, has been widely reported to participate in the process of coping with cold stress of plants. In this study, exogenous ALA promoted the growth of tomato plants and alleviated the appearance of purple tomato leaves under low-temperature stress. At the same time, exogenous ALA improved antioxidant enzyme activities, SlSOD gene expression, Fv/Fm, and proline contents and reduced H 2 O 2 contents, SlRBOH gene expression, relative electrical conductivity, and malondialdehyde contents to alleviate the damage caused by low temperature to tomato seedlings. Compared with low-temperature stress, spraying exogenous ALA before low-temperature stress could restore the indicators of photochemical quenching, actual photochemical efficiency, electron transport rate, and nonphotochemical quenching to normal. Exogenous ALA could increase the total contents of the xanthophyll cycle pool, the positive de-epoxidation rate of the xanthophyll cycle and improved the expression levels of key genes in the xanthophyll cycle under low-temperature stress. In addition, we found that exogenous ALA significantly enhanced the absorption of mineral nutrients, promoted the transfer and distribution of mineral nutrients to the leaves, and improved the expression levels of mineral nutrient absorption-related genes, which were all conducive to the improved adaptation of tomato seedlings under low-temperature stress. In summary, the application of exogenous ALA can increase tomato seedlings' tolerance to low-temperature stress by improving the xanthophyll cycle and the ability of the absorption of mineral nutrients in tomato seedlings. • Exogenous 5-aminolevulinic acid (ALA) significantly improved low temperature tolerance in tomatoes. • ALA could improve the xanthophyll cycle ability of tomato seedlings under low-temperature stress. • Exogenous ALA could improve nutrient uptake by tomato seedlings under low-temperature stress. • Exogenous ALA could promote the transformation of mineral nutrition to leaves under low-temperature stress. [ABSTRACT FROM AUTHOR]

• The effect of 5-aminolevulinic acid (ALA)-based photodynamic therapy (PDT), widely used in cancer therapy, on neuroblastoma is not well known. • Neuroblastoma cell lines were found to be susceptible to ALA-PDT. • ALA-PDT followed by ALA-photodynamic diagnosis navigation surgery might be a novel approach for the treatment of neuroblastoma. 5-Aminolevulinic acid (ALA)-based photodynamic therapy (PDT) is widely used in cancer therapy because of the tumor-specific accumulation of photosensitizing protoporphyrin IX (PpIX). We aimed to assess the susceptibility of human neuroblastoma cell lines to ALA-PDT and determine the mechanism of PDT. We used four human neuroblastoma cell lines (GOTO, NB9, IMR32, and NB1) and a gastric cancer cell line (MKN45) as a positive control. Cells were treated with increasing concentrations of ALA, and the ALA-induced production of PpIX in tumor cells was quantified using fluorescence spectrophotometry. PDT photocytotoxicity was measured by exposing the cells to a 630-nm irradiation for 10 min, and apoptotic cells stained with phosphatidylserine (PS) and propidium iodide (PI) were detected through flow cytometry. ALA cytotoxicity was not observed in any cell line. The intracellular concentration of PpIX increased in an ALA dose-dependent manner, and intracellular fluorescence of PpIX increased in a time-dependent manner. The viability of NB-1 cells treated with 250 μM 5-ALA rapidly decreased to 5%. Photocytotoxicity was observed in the following order: NB1, IMR32, NB-9, and GOTO. Photocytotoxicity was positively correlated with intracellular PpIX concentrations. PS+/PI- cells increased up to 21% after 12 h, and PS+/PI+ cells accounted for 35% of all cells after 24 h, which suggests that ALA-PDT induced apoptotic cell death. This study shows that neuroblastoma cell lines were susceptible to 5-ALA-PDT, resulting in persistent apoptotic cell death. N/A for basic study. [ABSTRACT FROM AUTHOR]

5-Aminolevulinic acid (5-ALA) is a non-protein amino acid widely used in agriculture, animal husbandry and medicine. Currently, microbial cell factories are a promising production pathway, but the lack of high-throughput fermentation strain screening tools often hinders the exploration of engineering strategies to increase cell factory yields. Here, mutant AC103-3H was screened from libraries of saturating mutants after response-specific engineering of the transcription factor AsnC of L-asparagine (Asn). Based on mutant AC103-3H, a whole-cell biosensor EAC103-3H with a specific response to 5-ALA was constructed, which has a linear dynamic detection range of 1–12 mM and a detection limit of 0.094 mM, and can be used for in situ screening of potential high-producing 5-ALA strains. With its support, overexpression of the C5 pathway genes using promoter engineering assistance resulted in a 4.78-fold enhancement of 5-ALA production in the engineered E. coli. This study provides an efficient strain screening tool for exploring approaches to improve the 5-ALA productivity of engineered strains. [ABSTRACT FROM AUTHOR]

In recent years, 5-aminolevulinic acid (5-ALA) has attracted significant interest due to its roles as a photodynamic prodrug and an antiviral agent. In this study, we present a new approach using aconitase A from Escherichia coli Nissle 1917 (EcNAcnA), renowned for its exceptional activity and conjunction with ALA synthase from Rhodobacter capsulatus (RcALAS) to enhance 5-ALA production in an engineered chassis. Expression of EcNAcnA and RcALAS via dual plasmids led to a 59 % increase in 5-ALA yield, reaching up to 6.645 g/L. Diverse 5-ALA production levels were observed with different combinations of promoters and replication origins for both genes. Subsequently, an all-in-one plasmid with a high copy number, designated as RcNN, was introduced into the genomic engineering RcI strain. This resulted in the production of 24.5 g/L 5-ALA with a productivity of 0.907 g/L/h in a bioreactor under pH control and glucose feeding over 27 h. To the best of our knowledge, this is the first study to enhance 5-ALA biosynthesis by applying a superior aconitase variant from E. coli Nissle 1917, which enhances isocitrate production in the tricarboxylic acid (TCA) cycle and alleviates reactive oxygen species (ROS), thereby promoting 5-ALA accumulation effectively. • EcNAcnA enhances the carbon flux to TCA cycle and reduce ROS in 5-ALA synthesis. • Coupling EcNAcnA and RcALAS keeps energy balance and triggers high-level 5-ALA. • The carbon source is crucial for 5-ALA rather than cell growth using dual genes. • The GroELS chaperone assists in protein folding but trades off in ALAS enzyme amount. • The optimal pH control and aeration applies in bioreactor maximizing 5-ALA production. [ABSTRACT FROM AUTHOR]

5-Aminolevulinic acid (5-ALA) is a prodrug of porphyrin IX (PpIX). Disadvantages of 5-ALA include poor stability, rapid elimination, poor bioavailability, and weak cell penetration, which greatly reduce the clinical effect of 5-ALA based photodynamic therapy (PDT). Presently, a novel targeting nanosystem was constructed using gold nanoparticles (AuNPs) as carriers loaded with a CSNIDARAC (CC9)-targeting peptide and 5-ALA via Au-sulphur and ionic bonds, respectively, and then wrapped in polylactic glycolic acid (PLGA) NPs via self-assembly to improve the antitumor effects and reduce the side effect. The successful preparation of ALA/CC9@ AuNPs-PLGA NPs was verified using ultraviolet-visible, Fourier transform infrared spectroscopy, and X-ray photoelectron spectroscopy. The analyses revealed good sphericity with a particle size of approximately140 nm, Zeta potential of 10.11 mV, and slow-controlled release characteristic in a weak acid environment. Confocal microscopy revealed targeting of NCL-H460 cells by NPs by actively internalising CC9 and avoiding the phagocytic action of RAW264.7 cells, and live fluorescence imaging revealed targeting of tumours in tumour-bearing mice. Compared to free 5-ALA, the nanosystem displayed amplified anticancer activity by increasing production of PpIX and reactive oxygen species to induce mitochondrial pathway apoptosis. Antitumor efficacy was consistently observed in three-dimensionally cultured cells as the loss of integrity of tumour balls. More potent anti-tumour efficacy was demonstrated in xenograft tumour models by decreased growth rate and increased tumour apoptosis. Histological analysis showed that this system was not toxic, with lowered liver toxicity of 5-ALA. Thus, ALA/CC9@AuNPs-PLGA NPs deliver 5-ALA via a carrier cascade, with excellent effects on tumour accumulation and PDT through passive enhanced permeability and retention action and active targeting. This innovative strategy for cancer therapy requires more clinical trials before being implemented. • A novel cascade-targeted delivery of 5-aminolevulinic acid nanosystem was constructed. • The nanosystem has a high drug delivery ability by active and passive tumour-targeting capabilities. • Endocytosis process is regulated by influencing ATP and vesicles, CLIC, and GP60 proteins. • Accumulation of protoporphyrin IX and reactive oxygen species amplified anticancer activity. • Antitumor efficacy is excellent in three-dimensional tumour spheres and xenograft tumour models. [ABSTRACT FROM AUTHOR]

As a precursor of protoporphyrin IX (PpIX), an endogenous pro-apoptotic and fluorescent molecule, 5-Aminolevulinic acid (5-ALA) has gained substantial attention for its potential in fluorescence-guided surgery as well as photodynamic therapy (PDT). Moreover, 5-ALA-PDT has been suggested as a promising chemo-radio sensitization therapy for various cancers. However, insufficient 5-ALA-induced PpIX fluorescence and the induction of multiple resistance mechanisms may hinder the 5-ALA-PDT clinical outcome. Reduced efficacy and resistance to 5-ALA-PDT can result from genomic alterations, tumor heterogeneity, hypoxia, activation of pathways related to cell surveillance, production of nitric oxide, and most importantly, deregulated 5-ALA transporter proteins and heme biosynthesis enzymes. Understanding the resistance regulatory mechanisms of 5-ALA-PDT may allow the development of effective personalized cancer therapy. Here, we described the mechanisms underlying resistance to 5-ALA-PTD across various tumor types and explored potential strategies to overcome this resistance. Furthermore, we discussed future approaches that may enhance the efficacy of treatments using 5-ALA-PDT. [Display omitted] [ABSTRACT FROM AUTHOR]

• 5-ALA-PDD for Pp IX fluorescence imaging of cholangiocarcinoma in the biliary tract. • Small-diameter flexible camera and light guide developed for per-oral cholangioscopy (POCS). • Pre-clinical study demonstrates devices efficiently detect Pp IX fluorescence. • This device combination is expected to enhance diagnostic accuracy in POCS. The diagnostic accuracy for cholangiocarcinoma (CCA) is inadequate, necessitating the exploration of novel diagnostic approaches. Protoporphyrin IX (Pp IX), a metabolic product of 5-aminolevulinic acid (5-ALA), emits red fluorescence upon blue light exposure. Because it accumulates selectively in cancer cells, photodynamic diagnosis using 5-ALA (5-ALA-PDD) has been integrated into clinical practice for diverse cancer types. Nevertheless, there is currently no device capable of capturing Pp IX-derived fluorescence for real-time 5-ALA-PDD within the biliary tract, largely due to challenges in device miniaturization. To investigate the feasibility of real-time 5ALA-PDD in CCA, we developed two essential components of the cholangioscopy system: a small-diameter flexible camera and a light guide for emitting blue light. We evaluated the detectability of Pp IX fluorescence using these devices in experimental gels and animal models. Our camera and light guide were smoothly inserted into the lumen of existing cholangioscopes. Incorporating a long-pass filter at the camera tip enabled efficient detection of red fluorescence without significantly impacting white-light observation. The integration of these devices facilitated clear visualization of red fluorescence from gels containing Pp IX at concentrations of 5 μM or higher. Additionally, when observing subcutaneous human CCA tumor models in nude mice treated with 5-ALA, we successfully demonstrated distinct red fluorescence from Pp IX accumulation in tumors compared to peritumoral subcutaneous areas. The integration of our device combination holds promise for real-time 5-ALA-PDD in human CCA, potentially enhancing the diagnostic accuracy for this complex condition. [ABSTRACT FROM AUTHOR]

• The photodynamic therapy was applied for the first time to treat dermatosis papulosa nigra(DPN), and its effectiveness and safety were preliminarily investigated. • This treatment method is expected to become a new and effective means of treating DPN. • Expand the treatment area of DPN and provide more effective treatment options. Dermatosis papulosa nigra (DPN) is a type of benign epidermal hyperplasia that affects the appearance of patients and poses a threat to their physical and mental health. Photodynamic therapy (PDT) has been shown to have the advantages of non-invasiveness, efficacy, and low recurrence in the treatment of skin disorders. However, no studies have been reported on the use of PDT for the treatment of DPN. Therefore, we used PDT for the first time to treat DPN and monitor its efficacy. Forty-five patients with DPN diagnosed at the outpatient clinic of Changzhou First People's Hospital were treated with 10 % 5-aminolevulinic acid (ALA) once a week for four consecutive weeks and followed for 3 months. After four treatment sessions, the cure rate among the 45 patients was 71.1 %, and the overall efficacy rate was 93.3 %. The most common adverse reactions were mild erythema, edema, and temporary pigmentation. PDT is expected to become a new and effective treatment for DPN. [ABSTRACT FROM AUTHOR]

• We evaluated the outcome of PDD-TURBT alone compared with white light TURBT with intravesical BCG therapy for intermediate- and high-risk NMIBC. • The difference in recurrence rate between PDD-TURBT alone and white light TURBT with intravesical BCG therapy tended to become more pronounced over time. • PDD-TURBT alone is insufficient to control the long-term recurrence of bladder cancer but can be effective in selected cases such as primary and single tumors. Although photodynamic-diagnosed transurethral resection of bladder cancer (PDD-TURBT) and Bacillus Calmette-Guérin (BCG) intravesical instillation are the two representative therapies for non-muscle invasive bladder cancer (NMIBC), no studies directly compare their efficacy. We evaluated the outcome of PDD-TURBT alone compared with white light TURBT with intravesical BCG therapy and analyzed the efficacy of both therapies depending on the characteristics of the tumors. We retrospectively analyzed intermediate- and high-risk NMIBC patients treated with PDD-TURBT alone (the PDD group) or white light TURBT with BCG therapy (the white light group) using propensity score matched analysis. In the propensity score matched cohort, the 1-, 2-, and 3-year recurrence-free survival rates for the PDD group were 77.6 %, 64.1 %, and 48.1 %, respectively, compared to 84.6 %, 75.1 %, and 75.1 % for the white light group (p = 0.44, 0.27, 0.17, respectively). The difference in recurrence rates between the two groups tended to become more pronounced over time, although there was no significant difference. In the univariate and multivariate analysis, recurrence, multiplicity, and tumor grade were the significant prognostic factors of recurrence in the PDD group (p = 0.010, 0.047, 0.048, respectively). Long-term RFS was similar in the PDD and white light groups when the population was limited to the primary and single tumors, suggesting that PDD-TURBT alone may be sufficient in this spectrum of patients. PDD-TURBT alone is insufficient to control the long-term recurrence of bladder cancer but can be effective in selected cases such as primary and single tumors. [ABSTRACT FROM AUTHOR]

• Modification of ALA-PDT is required to increase the phototoxicity efficiency. • ALA-PDT with hemin significantly increases PpIX accumulation and ROS production. • Hemin degradation into Fe2+ leads to iron accumulation, affects iron homeostasis. • PpIX, ROS, and Fe2+ accumulation triggers Fenton reaction, leading to cell death. Photodynamic therapy (PDT) has been utilized as a promising alternative cancer treatment due to its minimum invasiveness over the years. Exogenous 5-aminolevulinic acid (ALA) triggers protoporphyrin IX (PpIX) accumulation, which happens in cancer cells. However, certain types of cancer exhibit reduced effectiveness in the PpIX accumulation mechanism. This study aimed to determine the effect of ALA-PDT combination with hemin on gastric carcinoma TMK-1 cells. This study utilized TMK-1 gastric cancer cell line to evaluate PpIX, ROS, and Fe2+ accumulation following the administration of ALA, hemin, and a combination of ALA and hemin PDT. We also evaluate the mRNA expressions related to iron homeostasis and treatment impacts on cell viability. The co-addition of ALA and hemin PDT for 4 h of treatment resulted in a significant decrease in cell viability by up to 18 %. While ALA-PDT enhanced PpIX metabolism, the addition of hemin influenced both the production of reactive oxygen species (ROS) and cellular iron homeostasis by inducing Fe2+ accumulation and affecting mRNA levels of IRP, Tfr1, Ferritin, NFS1, and SDHB. These findings suggest that the addition of ALA and hemin enhances phototoxicity in TMK-1 cells. The combination of ALA and hemin with PDT induces cell death, evidenced by increased cytotoxicity properties such as PpIX and ROS, along with significant changes in TMK-1 gastric cancer iron homeostasis. Therefore, the combination of ALA and hemin could be one of the alternatives in photodynamic therapy for cancer in the future. [ABSTRACT FROM AUTHOR]

17α-Hydroxyprogesterone (17α-OH-PROG) is an important intermediate with a wide range of applications in the pharmaceutical industry. Strategies based on efficient electron transfer and cofactor regeneration were used for the production of 17α-OH-PROG. Here, CYP260A1, Fpr and Adx were expressed using a double plasmid system, resulting in higher biotransformation efficiency. Further optimization of reaction conditions and addition of polymyxin B increased the production of 17α-OH-PROG from 12.52 mg/L to 102.37 mg/L after 12 h of biotransformation. To avoid the addition of external 5-aminolevulinic acid (ALA) as a heme precursor for the P450 enzyme, a modified C5 pathway was introduced into the engineered strain, further reducing the overall process cost. The resulting whole-cell biocatalyst achieved the highest biotransformation yield of 17α-OH-PROG reported to date, offering a promising strategy for commercial application of P450 enzymes in industrial production of hydroxylated intermediates. [Display omitted] [ABSTRACT FROM AUTHOR]

Periodontitis, a chronic infectious disease leading to gingival atrophy and potential tooth loss through alveolar bone resorption, is closely linked to the oral microbiome. Fusobacterium nucleatum , known to facilitate late-stage bacterial colonization in the oral microbiome, plays a crucial role in the onset of periodontitis. Controlling F. nucleatum abundance is vital for preventing and treating periodontal disease. Photodynamic therapy combined with 5-aminolevulinic acid (ALA-PDT) has been reported to be bactericidal against Pseudomonas aeruginosa and Staphylococcus aureus. We aimed to investigate the bactericidal potential of ALA-PDT against F. nucleatum , which was evaluated by examining the impact of varying 5-ALA concentrations, culture time, and light intensity. After ALA-PDT treatment, DNA was extracted from interdental plaque samples collected from 10 volunteers and sequenced using the Illumina MiSeq platform. To further elucidate the bactericidal mechanism of ALA-PDT, porphyrins were extracted from F. nucleatum following cultivation with 5-ALA and subsequently analyzed using fluorescence spectra. ALA-PDT showed a significant bactericidal effect against F. nucleatum. Its bactericidal activity demonstrated a positive correlation with culture time and light intensity. Microbiota analysis revealed no significant alteration in α-diversity within the ALA-PDT group, although there was a noteworthy reduction in the proportion of the genus Fusobacterium. Furthermore, fluorescence spectral analysis indicated that F. nucleatum produced an excitable photosensitive substance following the addition of 5-ALA. Overall, if further studies confirm these results, this combined therapy could be an effective strategy for reducing the prevalence of periodontitis. • ALA-PDT has a significant bactericidal effect on F. nucleatum. • High-throughput analysis was used to examine the oral microbiota after ALA-PDT. • ALA-PDT can specifically remove F. nucleatum from the microbiota. • F. nucleatum generated uroporphyrin I after culture with 5-ALA. [ABSTRACT FROM AUTHOR]

• 5-ALA demonstrated growth inhibition in three vibrio species found in aquaculture. • Antimicrobial photodynamic therapy against vibrio spp. was more effective at pH 4.5. • 5-ALA exhibited antimicrobial activity under both light and dark conditions. • The optimal activity of 5-ALA under dark condition occurred with a 2 h exposure at pH 7. • Both crude and pure 5-ALA exhibited similar antimicrobial effects against vibrio species. Vibrio genus comprises a group of notorious pathogens that cause diseases in aquatic animals, leading to a reduction in seafood production. With climate change, Vibrio spp. pose an increasingly severe threat to the growing aquaculture industry, thereby affecting global food production. This study explored the use of 5-aminolevulinic acid (5-ALA) as a biocompatible antibacterial agent due to the inducibility of antibacterial properties by light and in vivo conversion to photosensitizing protoporphyrin IX (PPIX). 5-ALA were produced by recombinant E. coli was introduced as a photodynamic therapy against three Vibrio strains; namely, Vibrio parahaemolyticus (Vp), Vibrio alginolyticus (Va), and Vibrio damsela (Vd). The effect of 5-ALA on the three Vibrio species was investigated at a neutral pH, considering various factors such as reaction time and concentration under both light and dark conditions. The antimicrobial photodynamic therapy (aPDT) utilizing 0.5 % (w/v) 5-ALA was effective against Vibrio sp. only at pH < 5.5. In a neutral pH environment and without light exposure, antimicrobial activity was still observed against V. alginolyticus and Vibrio parahaemolyticus when the concentration of 5-ALA was increased to 1 % and 2 %, respectively. The bactericidal effect was only 39 % for Vibrio damsela, indicating a slower biosynthesis of PPIX. Nonetheless, this also suggests the potential of 5-ALA as an in vitro antibacterial agent under acidic conditions. The bactericidal effect of crude and pure 5-ALA was found to be similar, indicating the potential of using cost-effective crude extracts as aPDT agent. [ABSTRACT FROM AUTHOR]

• Fluorescein-sodium is comparable to 5-ALA and intraoperative MRI in terms of achieving gross-total resection of high-grade gliomas. • The use of intraoperative Fluorescein Sodium (vs. control) was associated with improved OS, and comparable rates of PFS as 5-ALA. • Fluorescein-sodium is an acceptable intraoperative agent that can be used effectively to guide surgical resection of high-grade gliomas. High grade gliomas (HGGs) are aggressive brain tumors associated with poor prognosis despite advances in surgical treatment and therapy. Navigated tumor resection has yielded improved outcomes for patients. We compare 5-ALA, fluorescein sodium (FS), and intraoperative MRI (IMRI) with no image guidance to determine the best intraoperative navigation method to maximize rates of gross total resection (GTR) and outcomes. A frequentist network meta -analysis was performed following standard PRISMA guidelines (PROSPERO registration CRD42021268659). Surface-under-the-cumulative ranking (SUCRA) analysis was executed to hierarchically rank modalities by the outcomes of interest. Heterogeneity was measured by the I2 statistic. Publication bias was assessed by funnel plots and the use of Egger's test. Statistical significance was determined by p < 0.05. Twenty-three studies were included for analysis with a total of 2,643 patients. Network meta -analysis comparing 5-ALA, IMRI, and FS was performed. The primary outcome assessed was the rate of GTR. Analysis revealed the superiority of all intraoperative navigation to control (no navigation). SUCRA analysis revealed the superiority of IMRI + 5-ALA, IMRI alone, followed by FS, and 5-ALA. Overall survival (OS) and progression free survival (PFS) were also examined. FS (vs. control) was associated with improved OS, while IMRI was associated with improved PFS (vs. control, FS, and 5-ALA). Intraoperative navigation using IMRI, FS, and 5-ALA lead to greater rates of GTR in HGGs. FS and 5-ALA also yielded improvement in OS and PFS. Further studies are needed to evaluate differences in survival benefit, operative duration, and cost. [ABSTRACT FROM AUTHOR]

• NA represents a severe manifestation of acne, and a subset of patients exhibit suboptimal response to isotretinoin. • Fire needle pretreatment ALA-PDT combined with low-dose isotretinoin in the treatment of refractory NA is effective and safe. • It's a clinical foundation for the management of refractory NA by fire needle pretreatment ALA-PDT combined with isotretinoin. Nodulocystic acne is a severe form of acne, which is commonly treated with oral isotretinoin, hormones, or antibiotics. However, drug therapy often has some side effects and poor compliance. Fire needle combined with 5-aminolevulinic acid photodynamic therapy (ALA-PDT) is a simple, effective, short-term treatment with few adverse reactions, which is expected to be an effective physiotherapy for nodulocystic acne. Moreover, the combination with isotretinoin can reduce the dosage of the drug, thereby reducing the side effects of isotretinoin. To evaluate the safety and efficacy of fire-needle pretreated ALA-PDT combined with low-dose isotretinoin in the treatment of severe refractory nodulocystic acne. This study reported 10 patients with refractory nodulocystic acne who received combined treatment. During the treatment period, all patients received a low dose of oral isotretinoin capsules daily. The acne lesions were pretreated with fire needle before ALA-PDT treatment. The number of acne lesions, including papules, pustules, and nodular cysts, was documented at weeks 0, 2, 4, 8, and 12 to assess the therapeutic efficacy. Concurrently, adverse reactions such as pain, pruritus, and pigmentation changes were recorded and evaluated throughout the treatment course. After combined treatment, all patients achieved good therapeutic effects, with an overall effective rate of 90 % at week 12. After treatment, skin lesions such as nodules, and cysts subsided significantly. The combination therapy has no serious adverse effects and has a favorable safety profile. Fire needle pretreatment ALA-PDT combined with low-dose isotretinoin is effective and safe in the treatment of severe refractory nodular cystic acne, which is worthy of clinical promotion and research. [ABSTRACT FROM AUTHOR]

Tomato is an important horticultural cash crop, and low-temperature stress has seriously affected the yield and quality of tomato. 5-Aminolevulinic acid (ALA) is widely used in agriculture as an efficient and harmless growth regulator. It is currently unclear whether exogenous ALA can cope with low-temperature stress by regulating tomato starch content and phenylalanine metabolism. In this study, exogenous ALA remarkably improved the low-temperature tolerance of tomato seedlings. RNA-sequencing results showed that exogenous ALA affected starch metabolism and phenylalanine metabolism in tomato seedling leaves under low-temperature stress. Subsequently, we used histochemical staining, observation of chloroplast microstructure, substance content determination, and qRT-PCR analysis to demonstrate that exogenous ALA could improve the low-temperature tolerance of tomato seedlings by regulating starch content and phenylalanine metabolism (SlPAL , SlPOD1 , and SlPOD2). Simultaneously, we found that exogenous ALA induced the expression of SlMYBs and SlWRKYs under low-temperature stress. In addition, dual luciferase, yeast one hybrid, and electrophoretic mobility shift assays indicate that SlMYB4 and SlMYB88 could regulate the expression of SlPOD2 in phenylalanine metabolism. We demonstrated that exogenous ALA could improve the low-temperature tolerance of tomato seedlings by regulating starch content and phenylalanine metabolism. • Exogenous 5-aminolevulinic acid (ALA) significantly improved low temperature tolerance in tomatoes. • ALA could regulating starch content and phenylalanine metabolism of tomato seedlings under low-temperature stress. • SlMYB4 and SlMYB88 could regulate the expression of SlPOD2 in phenylalanine metabolism. [ABSTRACT FROM AUTHOR]

Topical 5-aminolevulinic acid photodynamic therapy (PDT) and oral isotretinoin (ISO) are two effective treatments for moderate to severe acne vulgaris. Modified PDT (M-PDT) is associated with less pain, however, lacked a direct comparative evaluation with ISO. A multicenter, randomized clinical trial was conducted with participants randomly assigned to the PDT group or the ISO group and followed up to six months post-therapy. A total of 152 patients were allocated. The overall effective rates in the PDT group were significantly higher than that in the ISO group at one month, while the reverse was the case one month after treatment. No systemic side effects occurred in the PDT group while 70.67% of patients in ISO reported systemic side effects. M-PDT offers a more rapid onset of improvement, comparable overall efficacy, good tolerability, and comparable durability of response compared with ISO. [ABSTRACT FROM AUTHOR]

Photodynamic therapy (PDT) can be targeted toward different subcellular localizations and it is proposed that different subcellular targets vary in their sensitivity to photobiological damage. Since singlet oxygen (1O 2) has a very short lifetime with a limited diffusion length in cellular environments, measurement of cumulative 1O 2 luminescence is the most direct approach to compare the PDT sensitivity of mitochondria and plasma membrane. PDT-generated near-infrared 1O 2 luminescence at 1270 nm was measured together with cell viability for 5-aminolevulinic acid (ALA)-induced protoporphyrin IX (PpIX) and exogenous PpIX, at different incubation times. Confocal fluorescence microscopy indicated that ALA-induced PpIX (2 h) localized in the mitochondria, whereas exogenous PpIX (1 h) mainly localized to the plasma membrane. Cell viability was determined at several time points during PDT treatments using colony-forming assays, and the surviving fraction correlated well with cumulative 1O 2 luminescence counts from PpIX in mitochondria and plasmas membrane, respectively. The mitochondria are more sensitive than the plasma membrane by a factor of 1.7. The potential value of direct 1O 2 luminescence dosimetry for comparing the PDT sensitivity of different subcellular organelles was demonstrated, which could be useful for developing subcellular targeted-novel photosensitizers to enhance PDT efficiency. [ABSTRACT FROM AUTHOR]

• Chronic ulcers often present a therapeutic challenge in conventional medical approaches, with many cases exhibiting poor responsiveness. • Low-level ALA-PDT, characterized by the utilization of diminished concentrations of photosensitizers and reduced radiation doses, emerges as a promising therapeutic modality for managing chronic ulcers across a spectrum of complications. • Low-level ALA-PDT demonstrates efficacy in fostering complete healing of chronic ulcers and enhancing the overall quality of life for affected individuals. Chronic wounds refer to those that can't reconstruct anatomical and physical functional integrity, and are usually associated with signs of microbial infection. Current therapies include debridement and dressing change, local or systemic application of antibiotics, and medical dressing care, which are not ideal for the healing of chronic wounds. To explore the efficacy and safety of photodynamic therapy (ALA-PDT) for the treatment of chronic infectious wounds. ALA-PDT was used in ten patients with persistent wound infections and systemic complications who did not respond to conventional treatment. 5 % ALA solution was applied to the wound surface after debridement, incubated for 3 h with light protection, and then irradiated with red light for 20 min. This procedure was repeated every two weeks, and any adverse reactions were recorded. After the end of three treatments, the patients were followed up for 3 months. Patients who exhibit resistance to traditional therapies demonstrate a favorable therapeutic outcome with ALA-PDT, although complications may impede wound healing. All participants successfully underwent ALA-PDT treatment and subsequent monitoring, with 90 % achieving complete healing. Common adverse reactions to ALA-PDT encompass treatment-related pain, temporary erythema, and swelling, all of which are well-tolerated by patients without enduring severe consequences. ALA-PDT proves to be an efficacious intervention for managing chronic wounds, irrespective of the presence of localized infections or systemic complications. [ABSTRACT FROM AUTHOR]

• 5-ALA-conjugated bismuth oxide NPs are promising radiosensitizer and photosensitizer agents. • FA-5ALA-conjugated bismuth oxide NPs are promising radiosensitizer and photosensitizer agents. • There is significant synergistic effect of PDT and RT treatments in the presence of the conjugated NPs. Selective accumulation of photosensitizers into cancerous cells is one of the most important factors affecting photodynamic therapy (PDT) efficacy. 5-aminolevulinic acid (5-ALA) is the precursor of a strong photosensitizer, protoporphyrin-IX; but it has poor permeability into the cells. Folate receptors are overexpressed on the surface of many tumor cells. In the present study, folic acid (FA) and 5-ALA conjugated bismuth oxide nanoparticles were synthesized; and used in PDT, radiotherapy (RT), and concurrent PDT & RT against nasopharyngeal carcinoma (KB cell line). The KB cells were incubated with the synthesized nanoparticles (NPs) for 2 h; then illuminated using a custom-made LED lamp at the light dose of 26 J/cm2. Irradiation of the cells was carried out using X-ray 6 MV (2 Gy); and synergistic effect of the simultaneous RT and PDT treatments was evaluated using fractional product values. Efficacy of the treatments was determined using MTT and Caspase-3 enzyme activity assays. Targeting of folic acid receptors enables the selective endocytosis of the conjugated NPs. RT results in the presence of Bi 2 O 3 NPs showed a significant radiosensitizer potential of these NPs. Fractional product values of 1.49±0.05, 1.36±0.06, and 1.05±0.06 obtained in the presence of FA-5-ALA conjugated NPs, 5-ALA conjugated NPs, and in the absence of the NPs, respectively. Therefore, simultaneous RT and PDT in the presence of these conjugated NPs is superior to RT in the presence of the NPs. Simultaneous PDT and RT in the presence of FA-5-ALA conjugated bismuth oxide NPs can be introduced as a promising therapeutic approach in controlling KB cancer cells. [ABSTRACT FROM AUTHOR]

• 5-aminolevulinic acid photodynamic therapy (PDT) is non-invasive and causes no serious adverse reactions. • The efficacy of PDT, laser and PDT combined with laser pretreatment for low-grade vaginal intraepithelial neoplasia (VaIN1) was examined. • The three treatment are effective and have similar efficacy for high-risk HPV infection and VaIN1. • The addition of laser pretreatment does not increase the benefit of PDT treatment of VaIN1. • Older age, menopause, more times of pregnancies, and multiple vaginal lesions might affect HR-HPV regression. Vaginal intraepithelial neoplasia (VaIN) is a group of diseases of squamous epithelial dysplasia and carcinoma in situ occurring in the vagina, which is associated with high-risk human papillomavirus (HR-HPV) infection. To evaluate the efficacy and safety of Carbon dioxide (CO 2) laser, 5-aminolevulinic acid photodynamic therapy (PDT) and PDT combined with CO 2 laser pretreatment for VaIN1 with HR-HPV infection, and analyze the factors affecting the clearance of HR-HPV. Patients with HR-HPV infection and pathological diagnosis of VaIN1 and received laser or PDT or PDT combined with laser pretreatment were recruited. A total of 45 patients received one to three times CO 2 laser (laser Group), 15 patients received three times PDT (PDT Group) and 15 patients received CO 2 laser once and PDT three times (laser + PDT Group). HPV testing, cytology and colposcopy examinations at 3–6 months and 9–12 months after treatment were analyzed to assess the outcomes of the treatment. : There was no significant difference in regression rate of VaIN1 among the laser Group, the PDT Group and the laser + PDT Group (3–6 month follow-up: 57.78% vs 73.3% vs 80 %, 9–12 month follow-up: 68.89% vs 80% vs 86.67 %, P >0.05). HR-HPV remission rates were also similar in the three groups (3–6 month follow-up: 26.67% vs 46.67% vs 46.67 %, 9–12 month follow-up: 40 % in all groups, P >0.05). Compared to HR-HPV negative group, patients in the HR-HPV positive group were older and had more pregnancies. Menopause and multiple vaginal lesions were more common in the HR-HPV positive group. Adverse reactions were mild in the PDT Group. The laser Group and the laser + PDT Group had more adverse effects, such as increased vaginal secretion, vaginal bleeding, scarring and local pain. For patients with VaIN1 at risk of progression, ALA-PDT presents itself as a viable choice for those who are well-informed and can consent to its costs and benefits. The addition of CO 2 laser pretreatment may not increase the benefit of ALA-PDT treatment of VaIN1. Older age, menopause, more times of pregnancies, and multiple vaginal lesions might affect HR-HPV regression. [ABSTRACT FROM AUTHOR]

• ALA-PDT is an effective, safe, and alternative treatment for recurrent cervical HSIL after conization. • ALA-PDT has similar efficacy to surgery in patients with recurrent cervical HSIL after conization. • Compared with surgery, ALA-PDT protects the cervix from further damage. The study aimed to compare the clinical efficacy and safety of 5-aminolevulinic acid photodynamic therapy (ALA-PDT) and surgery in treating recurrent cervical high-grade squamous intraepithelial lesions (HSIL) after surgery due to precancerous lesions. A total of 41 patients with recurrent cervical HSIL after surgery for precancerous lesions were studied retrospectively. Patients underwent ALA-PDT or surgery and were followed up at 3, 6, 9 and 12 months and then every six months after that. Clinical data were collected and the efficacy and safety of the two treatment methods were compared. Of the 41 patients with recurrent cervical HSIL after conization, 15 cases received ALA-PDT and 26 received surgery. At the six-month follow-up, the lesions' complete remission (CR) rate was 93.33 % in ALA-PDT group and 88.46 % in the surgery group. The human papillomavirus (HPV) clearance rates were 66.67 % and 73.08 %, respectively. No significant differences concerning the lesions' CR rate and the HPV clearance rate were observed between the two groups (P >0.05). At the twelve-month follow-up, the HPV clearance rates were 80.00 % and 91.67 %. No significant differences concerning the HPV clearance rate were observed between the two groups (P >0.05). In the surgery group, the HPV clearance rate and the lesions' CR rate were lower in patients over 45 years of age (25.00% vs. 81.82 %, P = 0.031; 50.00% vs. 95.45 %, P = 0.052). During the follow-up, there was no significant difference in the recurrence rate between the two groups (P >0.05). In addition, none of the patients progressed. In women treated with ALA-PDT, there was no vaginal bleeding, and no harmful effects on the cervical organizational structure or functions compared to the surgery group, and two women delivered successfully after ALA-PDT treatment. The efficacy of ALA-PDT was similar to that of surgery in treating recurrent cervical HSIL following surgery, with fewer side effects. [ABSTRACT FROM AUTHOR]

5-Aminolevulinic acid (5-ALA) has been approved for clinical photodynamic therapy (PDT) due to its negligible photosensitive toxicity. However, the curative effect of 5-ALA is restricted by intracellular biotransformation inactivation of 5-ALA and potential DNA repair of tumor cells. Inspired by the crucial function of iron ions in 5-ALA transformation and DNA repair, a liposomal nanomedicine (MFLs@5-ALA/DFO) with intracellular iron ion regulation property was developed for boosting the PDT of 5-ALA, which was prepared by co-encapsulating 5-ALA and DFO (deferoxamine, a special iron chelator) into the membrane fusion liposomes (MFLs). MFLs@5-ALA/DFO showed an improved pharmaceutical behavior and rapidly fused with tumor cell membrane for 5-ALA and DFO co-delivery. MFLs@5-ALA/DFO could efficiently reduce iron ion, thus blocking the biotransformation of photosensitive protoporphyrin IX (PpIX) to heme, realizing significant accumulation of photosensitivity. Meanwhile, the activity of DNA repair enzyme was also inhibited with the reduction of iron ion, resulting in the aggravated DNA damage in tumor cells. Our findings showed MFLs@5-ALA/DFO had potential to be applied for enhanced PDT of 5-ALA. A liposomal nanomedicine (MFLs@5-ALA/DFO) was designed for boosting the PDT of 5-ALA through intracellular iron ion regulation. [Display omitted] [ABSTRACT FROM AUTHOR]

Sono-photodynamic therapy is a promising anticancer technique based on the combination of sonodynamic and photodynamic therapy to improve the cancer treatment effectiveness. This study was aimed at analyzing the effects of the sono-photodynamic (SPD) activity on protoporphyrin IX (PpIX) solution and PpIX-loaded rat liver. In vitro, PpIX 5 μM solutions were irradiated with light (635 nm, 30-50 mW/cm2), ultrasound (1 MHz, 1-2 W/cm2) and both. The PpIX absorption spectra recorded over exposure time revealed that the PpIX decay rate induced by SPD activity (combined irradiation) was approximately the sum of those induced by photodynamic and sonodynamic activity. In vivo, rats were intraperitoneally injected with 5-aminolevulinic acid at the dose of 500 mg/kg weight. After 3 h of injection, the PpIX-loaded livers were irradiated with light (635 nm, 180 ± 9 J/cm2), ultrasound (1.0 MHz, 770 ± 40 J/cm2) and both using a single probe capable of illuminating and sonicating the liver simultaneously. After 30 h, the liver damage induced by each protocol was analyzed histologically. It was found that a greater necrosis depth was induced by the SPD activity. These results suggest that the SPD activity could improve the PpIX decay rate and have greater scope than photodynamic or sonodynamic activity. Further studies should be performed to gain a better understanding of this protocol. [ABSTRACT FROM AUTHOR]

l A novel strain, PIECE, was obtained by integrating pdxY into an E. coli chromosome. l PIECE significantly increased both biomass and ALA production simultaneously. l Carbon flux was redirected to produce more glycine, OAA, and citric acid in PIECE. l ALAS activity was enhanced by co-expressing the chaperone GroELS in PIECE. l The highest ALA production reached 8.21 g/L using fed-batch fermentation. Recently, 5-aminolevulinic acid (ALA) has gained significant attention in research due to its extensive applications in agriculture, aquaculture, cosmetics, and photodynamic therapy for cancer. Microbial production of ALA is a green, high-efficient, and sustainable process. Herein, we have engineered a new strain of Escherichia coli called P dxY I ntegrative E. c oli E nhancer (PIECE). In this strain, the super salvage pathway was fine-tuned to enhance the regeneration of pyridoxal 5′-phosphate (PLP), on which the activity of ALA synthase (ALAS) is highly dependent. The PIECE strain, harboring ALA synthase (ALAS) from Rhodobacter capsulatus, produced 1.99 g/L ALA and 5.15 g/L biomass, that was a 4.33-fold increase in ALA production and a 1.84-fold increase in biomass compared to the parent strain. The metabolic flux analysis indicated that PIECE would redirect more carbon into the tricarboxylic acid (TCA) cycle and generate more glycine and glutamate, thereby increasing the ALA and biomass. Finally, the maximum ALA yield of 8.21 g/L and a productivity of 0.228 g/L/h were obtained by using fed-batch fermentation. The results showed that PIECE was not only feasible for high-level ALA production but also had significant potential to reprogram the carbon flux in the TCA cycle to produce value-added chemicals in the future. [Display omitted] [ABSTRACT FROM AUTHOR]

• A novel method combing microbial 5-ALA production with solid-state fermentation in feed processing was conducted. • Genetic modification and solid-state fermentation process optimization were conducted to improve 5-ALA production. • The engineered S. cerevisiae ScA3 was able to produce a high 5-ALA titer of 225.63 mg/kg dried materials. Over the past decade, 5-aminolevulinic acid (5-ALA) has been highlighted as a promising functional feed additive and immunomodulator for improving the general health, immune response, and resistance to disease of livestock and poultry. However, it is very costly to produce 5-ALA using conventional chemical synthesis methods. Classical microbial fermentation fulfills the criteria of environmental friendliness, but the unsatisfactory titers still hinder actual industrial production. This study aimed to develop a solid-state fermentation (SSF) process that can be used to efficiently enrich feed with 5-ALA at a low cost. First, the endogenous 5-ALA synthase was overexpressed in Saccharomyces cerevisiae via integrating a copy of HEM1 gene into the chromosome and introducing a multi-copy plasmid pRS416- HEM1 which constitutively overexpresses HEM1 gene. The resulting strain ScA3 was able to produce 63.82 mg/L 5-ALA in shake-flask fermentation. After process optimization, a titer of 225.63 mg/kg dry materials, exceeding the usual effective dosage reported in animal trials, was achieved within 48 h through SSF of 20 kg feed in a 90-L steel drum. To our knowledge, this is the first report on combining microbial 5-ALA production with SSF in feed processing, which will hopefully promote the application and popularization of 5-ALA in the feed industry. [ABSTRACT FROM AUTHOR]

Study Objective: To examine the diagnostic accuracy of hysteroscopic photodynamic diagnosis (PDD) using 5-aminolevulinic acid (5ALA) in patients with endometrial cancer and premalignant atypical endometrial hyperplasia.Design: A single-center, open-label, exploratory intervention study.Setting: University Hospital in Japan.Patients: Thirty-four patients who underwent hysteroscopic resection in the Department of Obstetrics and Gynecology at Keio University Hospital.Interventions: Patients were given 5ALA orally approximately 3 hours before surgery and underwent observation of the uterine cavity and endometrial biopsy using 5ALA-PDD during hysteroscopic resection. Specimens were diagnosed histopathologically and the diagnostic sensitivity and specificity of hysteroscopic 5ALA-PDD for malignancy in the uterine cavity was determined. Red (R), blue (B), and green (G) intensity values were determined from PDD images, and the relationships of histopathological diagnosis with these values were used to develop a model for objective diagnosis of uterine malignancy.Measurements and Main Results: Three patients were excluded from the study because of failure of the endoscope system. A total of 113 specimens were collected endoscopically. The sensitivity and specificity of 5ALA-PDD for diagnosis of malignancy in the uterine cavity were 93.8% and 51.9%, respectively. The R/B ratio in imaging analysis was highest in malignant lesions, followed by benign lesions and normal uterine tissue, with significant differences among these groups (p <.05). The R/B and G/B ratios were used in a formula for prediction of malignancy based on logistic regression and the area under the receiver operating characteristic curve for this formula was 0.838. At a formula cutoff value of 0.220, the sensitivity and specificity for diagnosis of malignant disease were 90.6% and 65.4%, respectively.Conclusion: To our knowledge, this is the first study of the diagnostic accuracy of 5ALA-PDD for malignancies in the uterine cavity. Hysteroscopic 5ALA-PDD had higher sensitivity and identifiability of lesions. These findings suggest that hysteroscopic 5ALA-PDD may be useful for diagnosis of minute lesions. [ABSTRACT FROM AUTHOR]

5-Aminolevulinic acid (5-ALA) is an unnatural amino acid and has been approved as a biodegradable, non-toxic pesticide and herbicide with applications in sustainable agriculture. 5-ALA can also be applied for cancer targeting via tumor localization and photodynamic therapy. Herein, we developed a feasible quantification, regulation and production method of 5-ALA in Escherichia coli is based on the chimera of 5-ALA synthetase from Rhodobacter sphaeroides (RshemA) and super-fold green fluorescent protein (sfGFP) under the control of dual promoters/double plasmids. 5-ALA production based on quantification with the reporter sfGFP was unsuccessfully for the RshemA-sfGFP fusion protein owing to a steric hindrance effect, but was effective using dual constitutive promoters (i.e., J23100 and P LacI) for RshemA and sfGFP independently. Moreover, a simple quantification method based on the linear relationship between 5-ALA concentration and the change in sfGFP intensity was calculated with the Hill equation according to the results of dual plasmids which composed of RshemA-threonine/homoserine exporter (RhtA) and the sensing plasmid pSU-T7-sfGFP. Compared with the conventional detection method for 5-ALA using Ehrlich's reagent, our proposed method is advantages in effectiveness, real-time detection, and outstanding sensitivity. Finally, the highest yield of 5-ALA was obtained in E. coli D2TT strain, reaching 2.46 g/L of 5-ALA produced in a 2.5-L baffle flask fermentation. Hence, this approach shows strong potential for improving 5-ALA production with appropriate regulation and detection based on the fluorescent signal. • Quantification of 5-ALA by sfGFP is successful in dual plasmids rather than used fusion protein. • sfGFP intensity with correlation of 5-ALA has demonstrated by Hill equation. • T7 and J23100 promoter showed the different regulation in genetic strains. • The highest yield of 5-ALA was produced by RshemA with RhtA under dual T7 promoter. [ABSTRACT FROM AUTHOR]

• Toolkits of different promoters and replication origins are evaluated in MR-1. • T7 RNA polymerase/promoter system is first proven in MR-1 for protein expression. • Addition of 0.5 mM IPTG at early lag-phase is the optimal condition for MR1::T7R. • Carbonic anhydrase capture of CO 2 reached 12,106 WAU/mg in genetic MR-1. • 4-times increment of 5-aminolevulinic acid production in genetic MR-1 via T7 system. Shewanella oneidensis MR-1 is a well-known electrogenic bacterium for its respiratory and extracellular electron transfer (EET) capability. However, the genetic toolkits, including promoters, replication origins and biological parts are still rarely used. In this study, constitutive promoters of pLacI, pJ23100, pJ23105, pJ23109, and pTet expressing super-folder green fluorescent protein (sfGFP) were verified in combination with replication origins of pBR322 and p15A. The optimal genetic module was obtained from the pLacI promoter and pBR322 origin, which the specific fluorescence intensity in the MR-1 reached 5518 a.u./g-DCW. T7RNA polymerase (T7RNAP) was also integrated into MR-1 chromosome (i.e., MR1::T7R) by homologous recombination to establish the T7 system. Thus, an expression vector was constructed under T7 promoter and a mobilization gene cluster, which was overexpressed on red fluorescence protein (RFP), carbonic anhydrase (CA), and heme-related proteins. The optimal condition for induction of isopropyl β- d -1-thiogalactopyranoside (IPTG) was determined by the RFP fluorescence intensity, which was 0.5 mM IPTG after 2 h incubation. Moreover, the carbon dioxide capture was enhanced with CAs which activity reached to 12,106 WAU/mg, while the productivity of valuable 5-aminolevuinic acid, a pro-drug for cancer therapy, increased by 3.96-folds with overexpression of HemD in MR1::T7R. The results proved the feasibility of the functional T7 system in Shewanella species. [ABSTRACT FROM AUTHOR]

Supramolecular peptide materials have attracted increasing attention due to their natural biological origin and versatile applications. However, it is often challenging to control and modulate the self-assembly of peptides (especially short peptides) for constructing hydrogels with tunable mechanical properties and adaptive injectability toward biomedical applications. Here, we report a supramolecular strategy for forming robust and injectable hydrogels based on the self-assembly of a rationally designed bola-dipeptide. The self-assembled hydrogels exhibit versatile functionalities, including flow under shear stress, good recovery properties, and easy encapsulation of hydrophilic prodrugs. The prodrug-loaded hydrogels show sustained release profiles, inhibited nontargeted leakage, and enhanced localized prodrug conversion, leading to highly efficient photodynamic tumor ablation. Hence, the supramolecular strategy is promising for the rational construction of injectable hydrogels toward targeted and sustained prodrug conversion and tumor therapy. Unlabelled Image • Biocompatible hydrogels were fabricated by self-assembly of a rationally designed bola-dipeptide. • The hydrogels show good thixotropy and recovery behaviors. • The hydrogels inhibited nontargeted leakage and enhanced conversion of a prodrug. • Photodynamic tumor ablation was realized using the prodrug-loaded hydrogels. [ABSTRACT FROM AUTHOR]

• 5-Aminolevulinic acid (5-ALA) causes postoperative increase in liver enzymes (PILE). • 5-ALA was given to patients with elevated preoperative liver enzymes (EPLE). • The incidence of PILE after 5-ALA administration was 45.5% in patients with EPLE. • Most PILEs were temporary and the incidence of severe PILE was low. • Administering 5-ALA is relatively safe even in patients with EPLE. The effect of 5-aminolevulinic acid (5-ALA) on a postoperative increase in liver enzymes (PILE) has not been clearly investigated in patients with elevated preoperative liver enzymes. The aim of this study was to evaluate the safety of 5-ALA in brain tumor patients with elevated preoperative liver enzymes by investigating the incidence, severity, and duration of PILE. This retrospective study was conducted in patients with a brain tumor who received 5-ALA preoperatively and had a preoperative increase in liver enzymes. Laboratory data on serum alanine transaminase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP), and total bilirubin (TB) levels were collected preoperatively and through postoperative day (POD) 30. The incidence and severity of PILE was evaluated based on the Common Terminology Criteria for Adverse Events. Among 134 patients, 61 (45.5%) developed 97 episodes (grade 1/2/3: 80/10/7) of PILE. Postoperative increases in ALT, AST, ALP, and TB were observed in 41 (30.6%), 33 (24.6%), 6 (4.5%), and 17 (12.7%) patients respectively. Five (3.7%) and two (1.5%) patients showed grade 3 increases in ALT and AST respectively. The maximum liver enzyme levels shown in 97 episodes resolved to preoperative baseline or normal (n = 70), grade 1 (n = 24), and grade 2 (n = 3) levels within POD 30. The incidence of PILE after 5-ALA-guided brain tumor surgery in patients with increased preoperative liver enzymes was 45.5%. However, most PILEs were temporary and the incidence of severe PILE was low, suggesting that administering 5-ALA is relatively safe even in patients with elevated preoperative liver enzymes. [ABSTRACT FROM AUTHOR]

• PpIX-based in vivo respirometry is a sensitive method. • In vivo and ex vivo respirometry provide different results. • Succinate attenuates LPS-induced decrease in oxygen consumption in vivo. Mitochondrial function has been predominantly measured ex vivo. Due to isolation and preservation procedures ex vivo measurements might misrepresent in vivo mitochondrial conditions. Direct measurement of in vivo mitochondrial oxygen tension (mitoPO 2) and oxygen disappearance rate (ODR) with the protoporphyrin IX‐triplet state lifetime technique (PpIX-TSLT) might increase our understanding of mitochondrial dysfunction in the pathophysiology of acute disease. LPS administration decreased mitochondrial respiration (ODR) in vivo but did not alter mitochondrial function as assessed with ex vivo techniques (high resolution respirometry and specific complex determinations). PpIX-TSLT measures in vivo mitoPO 2 and ODR and can be applied non-invasively at the skin. [ABSTRACT FROM AUTHOR]

This review analyses physical drug delivery enhancement technologies with a focus on improving UV damaged skin, actinic keratoses and non-melanoma skin cancer treatment. In recent years, physical drug delivery enhancement has been shown to enhance cosmeceutical and skin cancer treatment efficacy, but there are pros and cons to each approach which we discuss in detail. Mechanisms of action, clinical efficacy, experimental design, outcomes in academic publications, clinical trial reports and patents are explored to evaluate each technology with a critical, translation focused lens. We conclude that the commercial success of cosmeceutical applications, e.g. microneedles, will drive further innovation in this arena that will impact how actinic keratoses and non-melanoma skin cancers are clinically managed. Unlabelled Image [ABSTRACT FROM AUTHOR]

• Feline mammary tumors (FMT) showed good response to 5-ALA-PDT in vitro and in vivo. • In vitro evaluation of 5-ALA-PDT using 2D and 3D culture models of homologous tumor cells. • mRNA level assessment of the difference in inhibitory effect between 2D and 3D tumor cells in vitro experiment. Numerous studies have shown that photodynamic therapy (PDT) has a therapeutic effect on mammary tumor cells, with 5-aminolevulinic acid (5-ALA-HCL) being a commonly used photosensitizer for PDT. Feline mammary tumors (FMTs) are relatively common. However, the cytotoxic and antitumor effects of 5-ALA-PDT on FMTs have not been clarified. To this end, we evaluated the therapeutic effect of 5-ALA-PDT on FMTs through in vitro experiments using an FMT FKR cell line established for this study. We performed 5-ALA-PDT in 2D-cultured FKR-A (adherent cells) and 3D-cultured FKR-S (spheroid cells) cells and performed a series of studies to evaluate the cell viability and determine the protoporphyrin IX (PpIX) content in the cells as well as the expression levels of mRNAs associated with PpIX production and release. An in vivo study was performed to assess the effectiveness of 5-ALA-PDT. There was a significant difference in the concentration of PpIX in FMT cells under different incubation culture modes (2D versus 3D culture). The concentration of PpIX in FMT cells was correlated with the differences in cell culture (2D and 3D) as well as the expression levels of genes such as PEPT1, PEPT2, FECH, and HO-1. In the in vitro study, 5-ALA-PDT had a stronger inhibitory effect on 3D-cultured FKR-S cells, which resemble the internal environment of organisms more closely. We also observed a significant inhibitory effect of 5-ALA-PDT on FMT cells in vivo. To our knowledge, this is the first study on 5-ALA-PDT for FMTs under both 2D and 3D conditions. [ABSTRACT FROM AUTHOR]

• MGLSc patients with extensive glans or urethral involvement have poor outcomes and a high recurrence rate by conventional treatment. • ALA-PDT has a good therapeutic effect on patients with severe MGLSc with minimal side effects. • ALA-PDT provides a promising treatment option for severe MGLSc patients and is worth promoting in clinical practice. Male genital lichen sclerosus (MGLSc) typically impacts the external genitalia, resulting in balanitis, erectile pain, urination symptoms, and/or urinary retention. Urethral stricture develops in up to 20 % of these patients, which is usually found in the distal part of the urethra but can, in severe instances, impact the entire urethra and cause structural changes. Patients with skin lesions limited to the foreskin and partially extending to the glans can typically be cured by circumcision, but the recurrence rate of stricture is high when the glans or urethra is extensively involved. In the following case report, we describe a 45-year-old man with a history of MGLSc for 3 years and urethral stricture for 2 years, and these conditions remained untreated after circumcision. We emphasize that treatment with 5-aminolevulinic acid-induced photodynamic therapy (ALA-PDT) may further improve outcomes in such severe cases. [ABSTRACT FROM AUTHOR]