Search

5-Aminolevulinic acid (ALA), as a novel plant growth regulator, is a critical precursor for the biosynthesis of porphyrin compounds in all organisms. Many studies have reported that exogenous ALA treatment could improve fruit sweetness. However, the mechanism by which ALA promotes the increase in sugar content in fruit remains unclear. In this study, we found that ALA significantly promoted sucrose accumulation and SPS (sucrose phosphate synthase) activity in peach fruit. At 14, 28, 42, 50 and 60 days after ALA treatment, sucrose content of fruit was increased by 23%, 43%, 37%, 40% and 16%, respectively, compared with control treatment, and SPS enzyme activity was increased by 21%, 28%, 47%, 37% and 29%, respectively. Correlation analysis showed that the sucrose content of peach fruit under ALA treatment was significantly positively correlated with SPS activity. Subsequently, bioinformatics was used to identify SPS gene family members in peach fruit, and it was found that there were four members of the PpSPS gene family, distributed on chromosomes 1, 7 and 8, named PpSPS1, PpSPS2, PpSPS3 and PpSPS4, respectively. The results of qRT-PCR showed that PpSPS2 and PpSPS3 were highly expressed in response to ALA during fruit development, and the expression of PpSPS2 was positively correlated with SPS activity and sucrose accumulation in peach fruit. The results of tobacco subcellular localization showed that PpSPS2 was mainly distributed in the cytoplasm and nucleus, while PpSPS3 was mainly distributed in the nucleus. The results of this study will lay the foundation for further study on the functions of PpSPS and the regulation of sugar metabolism during the development and ripening of peach fruit by ALA.

Fusobacterium nucleatum (F. nucleatum), a key pathogen implicated in periodontal disease, contributes to oral biofilm maturation and is linked to development of systemic diseases like colorectal cancer and liver cirrhosis. Photodynamic therapy (PDT) combined with 5-aminolevulinic acid (5-ALA) treatment (ALA-PDT) selectively targets F. nucleatum by inducing porphyrin accumulation. The bactericidal effect of red light-based PDT on F. nucleatum has not been evaluated previously. This study investigates the effect of ALA-PDT using red light-emitting diode (LED) light on F. nucleatum subspecies and their porphyrin accumulation. F. nucleatum subspecies were cultured with varying concentrations of 5-ALA under anaerobic conditions. Porphyrin accumulation was measured via fluorescence spectroscopy, and colony-forming units were measured to determine bacterial viability post-treatment. Additionally, other subspecies responded well to 0.01% 5-ALA, and uroporphyrin I accumulation correlated with bacterial death, revealing optimal bactericidal conditions. These results suggest that optimizing light intensity and 5-ALA concentration can significantly enhance the therapeutic potential of ALA-PDT in oral healthcare.

Aminolevulinic acid (5-ALA) and dihydroporphyrin iron chelates (DHFe) play roles in plant growth regulation under normal and stressful conditions. However, definitive data on their roles in regulating chlorophyll synthesis in wheat are lacking. In this study, the optimal concentrations for foliar-spray application of 5-ALA and DHFe to promote wheat seedling growth and leaf chlorophyll content were assessed. The optimal concentrations of 5-ALA or DHFe (50 mg/L and 0.01 mg/L, respectively) were applied as foliar sprays to seedlings of wheat ‘Bainong 4199’ (high light efficient) and ‘Zhoumai No. 18’ (average light efficient) under different light intensities. Chlorophyll precursor contents and activities of three chlorophyll synthesis-related enzymes were altered, and the combined application of 5-ALA and DHFe more strongly increased the leaf chlorophyll content. 5-ALA significantly increased the contents of the chlorophyll precursors porphobilinogen, uroporphyrinogen III, protoporphyrin IX, and magnesium-protoporphyrin IX, and the activities of 5-amino-ketovalerate dehydratase and uroporphyrinogen III synthase. DHFe resulted in an increase in chlorophyll content but a significant decrease in chlorophyllase activity and protochlorophyllide content. 5-ALA promoted the synthesis of chlorophyll precursors by regulating the activities of 5-amino-ketovalerate dehydratase and uroporphyrinogen III synthase. DHFe decreased chlorophyllase activity, thereby slowing chlorophyll degradation and increasing the chlorophyll content.

5-Aminolevulinic acid (5-ALA), a non-proteinogenic amino acid, is an intermediate in the biosynthesis of heme and exerts antiviral effects against feline coronavirus (FCoV); however, the underlying mechanisms remain unclear. In the biosynthesis of heme, 5-ALA is condensed and converted to protoporphyrin IX (PpIX), which is then transformed into heme by the insertion of ferrous iron. Previous research has suggested that the metabolites generated during heme biosynthesis contribute to the antiviral effects of 5-ALA. Therefore, the present study investigated the in vitro mechanisms responsible for the antiviral effects of 5-ALA. The results obtained revealed that 5-ALA and PpIX both effectively reduced the viral titer in the supernatant of FCoV-infected fcwf-4 cells. Moreover, PpIX exerted virucidal effects against FCoV. We also confirmed that 5-ALA increased PpIX levels in cells. While hemin induced heme oxygenase-1 gene expression, it did not reduce the viral titer in the supernatant. Sodium ferrous citrate decreased PpIX levels and suppressed the antiviral effects of 5-ALA. Collectively, these results suggest that the antiviral effects of 5-ALA against FCoV are dependent on PpIX.

Background: Minimally-invasive 5-aminolevulinic acid photodynamic therapy (ALA-PDT) is used for treating cervical high-grade squamous intraepithelial lesions (HSIL/CIN2). The purpose of this study was to analyze the factors affecting the efficacy of ALA-PDT in the treatment of cervical HSIL/CIN2 in order to guide physicians in making appropriate treatment decisions. Methods: A retrospective study including 69 female patients with pathologically diagnosed HSIL/CIN2 was conducted. Patients were given six doses of 20% ALA-PDT at 7–14-day intervals. Cytology, HPV testing, colposcopy, and pathology were performed before treatment and at 6-month follow-up after treatment to assess efficacy. The main outcome of this study was the regression of HSIL/CIN2 and the clearance of high-risk HPV (hrHPV) infection after ALA-PDT treatment. Clinicopathological characteristics were collected to analyze the factors affecting the effectiveness of ALA-PDT treatment for HSIL/CIN2. Results: Between the successful and failed lesion regression group, there was a significant difference in sleeping disorders (p < 0.05). Between the successful and failed hrHPV clearance group, no statistically significant factors were found. With sensitivity values of 0.556 and 0.778, respectively, multivariate analysis showed that current smoking and sleeping disorders were independent prognostics of failure in lesion regression after ALA-PDT treatment. Conclusions: Smoking and sleep disorders were independent risk factors for failure in HSIL/CIN2 regression following ALA-PDT, suggesting the need for careful consideration of ALA-PDT for patients with these conditions.

Oral squamous-cell and pancreatic carcinomas are aggressive cancers with a poor outcome. Photodynamic therapy (PDT) consists of the use of photosensitizer-induced cell and tissue damage that is activated by exposure to visible light. PDT selectively acts on cancer cells, which have an accumulation of photosensitizer superior to that of the normal surrounding tissues. 5-aminolevulinic acid (5-ALA) induces the production of protoporphyrin IX (PpIX), an endogenous photosensitizer activated in PDT. This study aimed to test the effect of a new gel containing 5% v/v 5-ALA (ALAD-PDT) on human oral CAL-27 and pancreatic CAPAN-2 cancer cell lines. The cell lines were incubated in low concentrations of ALAD-PDT (0.05%, 0.10%, 0.20%, 0.40%, 0.75%, 1.0%) for 4 h or 8 h, and then irradiated for 7 min with 630 nm RED light. The cytotoxic effects of ALAD-PDT were measured using the MTS assay. Apoptosis, cell cycle, and ROS assays were performed using flow cytometry. PpIX accumulation was measured using a spectrofluorometer after 10 min and 24 and 48 h of treatment. The viability was extremely reduced at all concentrations, at 4 h for CAPAN-2 and at 8 h for CAL-27. ALAD-PDT induced marked apoptosis rates in both oral and pancreatic cancer cells. Elevated ROS production and appreciable levels of PpIX were detected in both cell lines. The use of ALA-PDT as a topical or intralesional therapy would permit the use of very low doses to achieve effective results and minimize side effects. ALAD-PDT has the potential to play a significant role in complex oral and pancreatic anticancer therapies.

Extracorporeal photopheresis (ECP) is a therapeutic modality used for T-cell-mediated disorders. This approach involves exposing isolated white blood cells to photoactivatable 8-methoxypsoralen (8-MOP) and UVA light, aiming to induce apoptosis in T-cells and thereby modulate immune responses. However, conventional 8-MOP-ECP lacks cell selectivity, killing both healthy and diseased cells, and has shown limited treatment efficacy. An alternative approach under investigation involves the use of 5-aminolevulinic acid (ALA) in conjunction with light, referred to as ALA-based photodynamic therapy. Our previous ex vivo studies suggest that ALA-ECP exhibits greater selectivity and efficiency in killing T-cells derived from patients with T-cell-mediated disorders compared to those treated with 8-MOP-ECP. We have conducted a clinical phase I–(II) study evaluating ALA-ECP safety and tolerability in cutaneous T-cell lymphoma (CTCL). Here, 20 ALA-ECP treatments were administered to one CTCL patient, revealing no significant changes in vital signs. Two adverse events were reported; both evaluated by the Internal Safety Review Committee as non-serious. In addition, five conceivable events with mainly mild symptoms took place. During the study period, a 53% reduction in skin involvement and a 50% reduction in pruritus was observed. In conclusion, the results indicate that ALA-ECP treatment is safe and well tolerated.

Persistent HPV infections may cause cervical and vaginal intraepithelial neoplasia (CIN and VaIN). Traditional methods might destroy the structure and function of the cervix. 5-aminolevulinic acid photodynamic therapy (ALA-PDT) is a non-invasive targeted therapy. This study aims to evaluate the efficacy and safety of ALA-PDT for CIN and VaIN and the clearance of HPV. A retrospective study of 303 patients who confirmed CIN or VaIN and received ALA-PDT was conducted. All the patients were followed up at six and twelve months after treatment and then annually thereafter. The effect was evaluated through HPV genotyping, a cytology test, and colposcopy-directed biopsy if necessary. After ALA-PDT, the remission rates for CIN 2, CIN 3, VaIN 2, and VaIN 3 were 90.6%, 88.5%, 87.3%, and 77.8%. For CIN 1, the remission rate at the six-month follow-up was 93.1%. The total HPV clearance rates were 72.5% at the six-month follow-up and 85.7% at the 12-month follow-up. The most common adverse event was vaginal discharge. No severe adverse effect was observed. ALA-PDT is an effective and safe treatment for all grades of CIN and VaIN and is helpful in clearing HPV with minimal side effects. This treatment may not influence fertility and delivery.

Using beneficial microorganisms, such as purple non-sulfur bacteria (PNSB), has shown enormous potential for improving plant growth and agricultural production. However, the full extent of their benefits and interactions with agricultural practices is yet to be fully understood. The present study aimed to investigate the use of PNSB in crop rotation practice, focusing on its impact on rice growth and yield. The experiment was conducted over two rice cropping seasons, with djulis grown between the rice as a rotation crop. The study shows that PNSB treatment increased the concentration of 5-aminolevulinic acid (5-ALA) in plants, indicating enhanced photosynthesis. Moreover, when combined with crop rotation, PNSB remarkably improved soil fertility. These combined benefits resulted in substantial increases in tiller numbers (163%), leaf chlorophyll content (13%), and lodging resistance (66%), compared to the untreated plants. The combined treatment also resulted in higher productive tillers per hill (112%), average grain per hill (65%), and grain fertility (26%). This led to increased grain yield (65%), shoot dry weight (15%), and harvest index (37%). The findings clearly suggest that the incorporation of PNSB in crop rotation strategies can significantly augment the growth and yield of rice crops. These insights, pivotal for sustainable rice cultivation, hold the potential to simultaneously tackle the pressing issues of global food security and climate change.

Cutaneous alternariosis is a rare condition, more frequently presented in immunocompromised patients, which usually requires long courses of systemic antifungals that may interact with other medications. The presented series shows three cases of cutaneous alternariosis in immunocompromised patients and organ transplant recipients that were successfully treated with photodynamic therapy and oral antifungals, allowing a reduction in the systemic treatment duration and therefore decreasing the risk of side effects and drug interactions.

Fusobacterium nucleatum ( F. nucleatum ), a key pathogen implicated in periodontal disease, contributes to oral biofilm maturation and is linked to development of systemic diseases like colorectal cancer and liver cirrhosis. Photodynamic therapy (PDT) combined with 5-aminolevulinic acid (5-ALA) treatment (ALA-PDT) selectively targets F. nucleatum by inducing porphyrin accumulation. The bactericidal effect of red light-based PDT on F. nucleatum has not been evaluated previously. This study investigates the effect of ALA-PDT using red light-emitting diode (LED) light on F. nucleatum subspecies and their porphyrin accumulation. F. nucleatum subspecies were cultured with varying concentrations of 5-ALA under anaerobic conditions. Porphyrin accumulation was measured via fluorescence spectroscopy, and colony-forming units were measured to determine bacterial viability post-treatment. Additionally, other subspecies responded well to 0.01% 5-ALA, and uroporphyrin I accumulation correlated with bacterial death, revealing optimal bactericidal conditions. These results suggest that optimizing light intensity and 5-ALA concentration can significantly enhance the therapeutic potential of ALA-PDT in oral healthcare.

Background: Minimally-invasive 5-aminolevulinic acid photodynamic therapy (ALA-PDT) is used for treating cervical high-grade squamous intraepithelial lesions (HSIL/CIN2). The purpose of this study was to analyze the factors affecting the efficacy of ALA-PDT in the treatment of cervical HSIL/CIN2 in order to guide physicians in making appropriate treatment decisions., Methods: A retrospective study including 69 female patients with pathologically diagnosed HSIL/CIN2 was conducted. Patients were given six doses of 20% ALA-PDT at 7-14-day intervals. Cytology, HPV testing, colposcopy, and pathology were performed before treatment and at 6-month follow-up after treatment to assess efficacy. The main outcome of this study was the regression of HSIL/CIN2 and the clearance of high-risk HPV (hrHPV) infection after ALA-PDT treatment. Clinicopathological characteristics were collected to analyze the factors affecting the effectiveness of ALA-PDT treatment for HSIL/CIN2., Results: Between the successful and failed lesion regression group, there was a significant difference in sleeping disorders ( p < 0.05). Between the successful and failed hrHPV clearance group, no statistically significant factors were found. With sensitivity values of 0.556 and 0.778, respectively, multivariate analysis showed that current smoking and sleeping disorders were independent prognostics of failure in lesion regression after ALA-PDT treatment., Conclusions: Smoking and sleep disorders were independent risk factors for failure in HSIL/CIN2 regression following ALA-PDT, suggesting the need for careful consideration of ALA-PDT for patients with these conditions.

Saline soils negatively affect and cause serious problems for rice cultivation. This study aimed to evaluate the efficacy of the purple nonsulfur bacteria (PNSB) capable of secreting 5-aminolevulinic acid (ALA) to reduce soil salinity, improve soil fertility, and enhance rice growth and yield. A two-factorial experiment was conducted in a randomized complete block design with four replications. Factor one was the salinity of the irrigated water, and factor two was the supplementation of the ALA-producing PNSB. The results indicated that watering with saline water above 3‰ led to decreases in plant growth and rice yield compared to the treatments watered with tap water. Application of either an individual strain or the mixture of W01, W14, and W22 ameliorated soil properties and increased total NPK uptake, whereas treatments supplied with the mixed strains reduced total Na uptake (9.50 mg Na pot−1). Supplying the W01, W14, and W22 strains individually or in a mixture enhanced the plant height by 3.51–5.45% and rice grain yield by 14.7–26.2%, compared with those of the control treatment. From the study, the combination of the L. sphaeroides W01, W14, and W22 strains is promising for application in saline or salt-contaminated regions to aid the damages caused by salinity on cultivars there, especially rice. Furthermore, this is a biological approach to ease an environmental problem and improve crop performance, which is supposed to be a trend in the sustainable agriculture.

Recurrent glioblastoma multiforme (GBM) is largely attributed to peritumoral infiltration of tumor cells. As higher CD44 expression in the tumor periphery correlates with higher risk of GBM invasion, the present study analyzed the relationship between CD44 expression and magnetic resonance imaging (MRI)-based invasiveness of GBM on a large scale. We also quantitatively evaluated GBM invasion using 5-aminolevulinic acid (5-ALA) spectroscopy to investigate the relationship between CD44 expression and tumor invasiveness as evaluated by intraoperative 5-ALA intensity. Based on MRI, GBM was classified as high-invasive type in 28 patients and low-invasive type in 22 patients. High-invasive type expressed CD44 at a significantly higher level than low-invasive type and was associated with worse survival. To quantitatively analyze GBM invasiveness, the relationship between tumor density in the peritumoral area and the spectroscopic intensity of 5-ALA was investigated. Spectroscopy showed that the 5-ALA intensity of infiltrating tumor cells correlated with tumor density as represented by the Ki-67 staining index. No significant correlation between CD44 and degree of 5-ALA-based invasiveness of GBM was found, but invasiveness of GBM as evaluated by 5-ALA matched the classification from MRI in all except one case, indicating that CD44 expression at the GBM periphery could provide a reliable biomarker for invasiveness in GBM.

Heme b, which is characterized by a ferrous ion and a porphyrin macrocycle, acts as a prosthetic group for many enzymes and contributes to various physiological processes. Consequently, it has wide applications in medicine, food, chemical production, and other burgeoning fields. Due to the shortcomings of chemical syntheses and bio-extraction techniques, alternative biotechnological methods have drawn increasing attention. In this review, we provide the first systematic summary of the progress in the microbial synthesis of heme b. Three different pathways are described in detail, and the metabolic engineering strategies for the biosynthesis of heme b via the protoporphyrin-dependent and coproporphyrin-dependent pathways are highlighted. The UV spectrophotometric detection of heme b is gradually being replaced by newly developed detection methods, such as HPLC and biosensors, and for the first time, this review summarizes the methods used in recent years. Finally, we discuss the future prospects, with an emphasis on the potential strategies for improving the biosynthesis of heme b and understanding the regulatory mechanisms for building efficient microbial cell factories.

This study aimed to investigate the anti-inflammatory effect of 5-aminolevulinic acid (5-ALA) on endotoxin-induced uveitis (EIU) in rats. EIU was induced in male Sprague Dawley rats by the subcutaneous injection of lipopolysaccharide (LPS). During LPS injection, 5-ALA diluted with saline was administered via gastric gavage. After 24 h, clinical scores were assessed after which aqueous humor (AqH) samples were obtained. The number of infiltrating cells, protein concentration, and levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), nitric oxide (NO), and prostaglandin E2 (PGE2) in AqH were measured. For histological examination, both eyes of some rats were enucleated. In vitro, a mouse macrophage cell line (RAW264.7 cells) was stimulated by LPS with or without 5-ALA. Western blot was used to analyze the expression of inducible NO synthase (iNOS) and cyclooxygenase-2. 5-ALA suppressed the EIU clinical scores, infiltrating cell number, and protein concentration while improving the histopathologic scores. In particular, 100 mg/kg 5-ALA reduced the concentrations of NO, PGE2, TNF-α, and IL-6 in AqH, similar to 1 mg/kg prednisolone. In addition, 5-ALA suppressed iNOS upregulation in LPS-stimulated RAW264.7 cells. Therefore, 5-ALA has an anti-inflammatory effect on EIU through the inhibition of the upregulation of inflammatory mediators.

5-Aminolevulinic acid (5-ALA) has recently received much attention for its wide applications in medicine and agriculture. In this study, we investigated the effect of NCgl0580 in Corynebacterium glutamicum on 5-ALA biosynthesis as well as its possible mechanism. It was found that the overexpression of NCgl0580 increased 5-ALA production by approximately 53.3%. Interestingly, the knockout of this gene led to an even more significant 2.49-fold increase in 5-ALA production. According to transcriptome analysis and functional validation of phenotype-related targets, the deletion of NCgl0580 brought about considerable changes in the transcript levels of genes involved in central carbon metabolism, leading to fluxes redistribution toward the 5-ALA precursor succinyl-CoA as well as ATP-binding cassette (ABC) transporters affecting 5-ALA biosynthesis. In particular, the positive effects of enhanced sugar transport (by overexpressing NCgl1445 and iolT1), glycolysis (by overexpressing pyk2), iron uptake (by overexpressing afuABC), and phosphate uptake (by overexpressing pstSCAB and ugpQ) on 5-ALA biosynthesis were demonstrated for the first time. Thus, the transcriptional mechanism underlying the effect of NCgl0580 deletion on 5-ALA biosynthesis was elucidated, providing new strategies to regulate the metabolic network of C. glutamicum to achieve a further increase in 5-ALA production.

Guided bone regeneration (GBR) comprehends the application of membranes to drive bone healing and to exclude non-osteogenic tissues from interfering with bone regeneration. However, the membranes may be exposed to bacterial attack, with the risk of failure of the GBR. Recently, an antibacterial photodynamic protocol (ALAD-PDT) based on a gel with 5% 5-aminolevulinic acid incubated for 45 min and irradiated for 7 min by a LED light at 630 nm, also showed a pro-proliferative effect on human fibroblasts and osteoblasts. The present study hypothesized that the functionalization of a porcine cortical membrane (soft-curved lamina, OsteoBiol) with ALAD-PDT might promote its osteoconductive properties. TEST 1 aimed to verify the response of osteoblasts seeded on lamina with respect to the plate surface (CTRL). TEST 2 aimed to investigate the effects of ALAD-PDT on the osteoblasts cultured on the lamina. SEM analyses were performed to study the topographical characteristics of the membrane surface, the adhesion, and the morphology of cells at 3 days. The viability was assessed at 3 days, the ALP activity at 7 days, and calcium deposition at 14 days. Results showed the porous surface of the lamina and the increase in cell attachment of osteoblasts with respect to controls. The proliferation, the ALP, and bone mineralization activity of osteoblasts seeded on lamina resulted in being significantly higher (p < 0.0001) than controls. Results also showed an additional significative enhancement (p < 0.0001) in the proliferative rate in ALP and calcium deposition after applying ALAD-PDT. In conclusion, the functionalization of the cortical membranes cultured with osteoblasts with the ALAD-PDT improved their osteoconductive properties.

Manganese (Mn) is an essential nutrient in most organisms. Establishing an effective regulatory system of Mn absorption is important for sustainable crop development. In this study, we selected sunflower as the model plant to explore the effects of 5-aminolevulinic acid (ALA) or salicylic acid (SA) combined with citric acid (CA) on Mn absorption. Six-leaf-old sunflower plants were exposed to 0.8 g kg−1 Mn for one week and then treated with chelating agents, i.e., CA (10 mmol kg−1), and different concentrations of ALA and SA for one week. The results showed that Mn-treated plants had significantly increased H2O2, O2− and MDA contents in leaves compared with the control. Under the Mn + CA treatment, ALA or SA2 significantly activated the antioxidant defense system by increasing SOD, POD and CAT activities in leaves. Moreover, the application of CA significantly increased the Mn uptake in sunflower roots compared with Mn treatment alone; however, did not accelerate the translocation efficiency of Mn from sunflower roots to shoots. Moreover, ultrastructural and RT-qPCR results further demonstrated that ALA/SA could recover the adverse impact of excessive Mn accumulation in sunflowers. Like a pump, ALA/SA regulated the translocation efficiency and promoted the transportation of Mn from roots to shoots. This study provides insights into the promotive role of ALA/SA combined with CA on sunflower growth by regulating Mn absorption, which would be beneficial for regulating Mn absorption in soil with an Mn deficit.

Background: In this ex vivo study, the aim was to evaluate the effects of ALAD and red light on Enterococcus faecalis in infected root canals using a special intracanal fiber. Methods: A total of 70 extracted, single-rooted teeth were used. The teeth were decoronated at the length of the roots to approximately 15 mm and then instrumented. The apical foramen was sealed by composite resin, and the root canals were infected with a pure culture of E. faecalis ATCC 29212 for eight days at 37 °C. Following the contamination period, the roots were divided into seven groups, including the positive and negative control groups, and treated as follows: ALAD 45 min; red light activation 7 min; ALAD 45 min and red-light activation 7 min; sodium hypochlorite 2.5% 15 min; sodium hypochlorite 1% 15 min. The samples were taken by three sterile paper points, transferred to tubes containing 1 mL of PBS, and immediately processed for the number of colony-forming units and the cell viability by using live/dead. Results: The best treatment is obtained with 2.5% NaOCl. Except for ALAD + red light vs. 1% NaOCl, a statistically significant difference is recorded for all treatments. The combination of 2.5% NaOCl and ALAD + 7 min irradiation produces an evident killing effect on the E. faecalis cells. On the other hand, 1% NaOCl is ineffective for the viability action, with 25% of dead cells stained in red. Conclusions: This ex vivo study shows that ALAD gel with light irradiation is an efficacious protocol that exerts a potent antibacterial activity against E. faecalis in infected root canals.

Non-invasive detection of polarized macrophages in tumors is an urgent task in terms of combined antitumor therapy. By analyzing the fluorescence lifetime of the metabolic cofactors—the reduced form of nicotinamide adenine dinucleotide (NADH) and flavins—differences in cellular metabolism of normal tissue, tumor, inflammatory and anti-inflammatory macrophages were demonstrated. In this work we studied changes in the polarization of macrophages obtained from THP-1 monocytes in response to photodynamic therapy with 5-aminolevulinic acid (ALA–PDT). Moderate ALA–PDT in vitro led to changes in M0 macrophages metabolism towards M1 polarization, wherein M1 and M2 macrophages died and were replaced by non-polarized cells. The interstitial distribution of polarized macrophages after ALA–PDT was studied in a mouse tumor model of grafted Lewis lung carcinoma. In response to ALA–PDT, there was an increase in the inflammatory macrophages fraction in the tumor node. Metabolic fluorescence-lifetime imaging microscopy (FLIM) was performed for macrophages in vitro and for tumor cryosections. It was shown that analysis of phasor diagrams for the NADH, flavins, and 5-ALA-induced protoporphyrin IX (PpIX) fluorescence lifetime helps to determine the change in metabolism in response to different modes of PDT at the cellular and tissue levels. These data can be used for post-surgery tissue inspection.

This study focused on the physiological regulation and mechanism of exogenous 5-aminolevulinic acid (5-ALA) in the late growth of P. heterophylla. In the middle of May, different concentrations of 5-ALA (0, 10, 20, 50 mg·L−1) were sprayed on the leaves. The effects of 5-ALA on tuberous root growth, antioxidant enzyme system, gas exchange, photosynthetic pigment contents and photosynthetic characteristics were measured from 23 May to 13 June. A concentration of 20 mg·L−1 of 5-ALA led to a significant increase in the yield of fresh root and biomass allocation at 38.12% and 25.07%, respectively, in comparation with the control (0 mg·L−1). The moderate concentration of 5-ALA statistically stimulated antioxidation activities. 5-ALA treatment enhanced photosynthetic activity and reduced photodamage. Compared to the control, there were increases in the chlorophyll fluorescence parameters of P. heterophylla under 5-ALA treatment. Moreover, 20 mg·L−1 of 5-ALA significantly changed the kinetic parameters of fluorescence. It enhanced the light absorption and distribution efficiency of PSII and the activities of leaves, resulting in alleviating photoinhibition by the excess excitation energy. The correlation indicated that there was a significant positive correlation between the yield of tuberous roots and biomass allocation, Pn and catalase (CAT), and a negative correlation between the yield of tuberous roots and malondialdehyde (MDA). The appropriate 5-ALA concentration in the late growth stage of P. heterophylla effectively enhanced the net photosynthetic capacity, mainly resulting from the enhancement of PSII photochemical activity to promote the increases in excitation energy absorption, capture and electron transfer efficiency of the leaves. Finally, 5-ALA treatment can increase the photochemical activity of PSII in the whole leaf and ultimately delay the senescence of P. heterophylla.

The present study aims to discuss the main factors involving the use of 5-aminolevulinic acid together with red LED light and its application in endodontic treatment through a narrative review and a case report. Persistence of microorganisms remaining on chemical-mechanical preparation or intracanal dressing is reported as the leading cause of failure in endodontics. Photodynamic therapy has become a promising antimicrobial strategy as an aid to endodontic treatment. Being easy and quick to apply, it can be used both in a single session and in several sessions, as well as not allowing forms of microbial resistance. 5-aminolevulinic acid in combination with red LED light has recently been studied in many branches of medicine, with good results against numerous types of bacteria including Enterococuss faecalis. The case report showed how bacterial count of CFU decreased by half (210 CFU/mL), after 45 min of irrigation with a gel containing 5% of 5-aminolevulinic acid compared to the sample before irrigation (420 CFU/mL). The subsequent irradiation of red LED light for 7 min, the bacterial count was equal to 0. Thus, it is concluded that the use of 5-aminolevulinic acid together with red LED light is effective in endodontic treatment.

Using female Sprague–Dawley (SD) rats as a model, the current study aimed to investigate whether feeding 5-aminolevulinic acid (5-ALA) to female SD rats during gestation and lactation can affect the iron status of weaned rats and provide new ideas for the iron supplementation of piglets. A total of 27 pregnant SD rats were randomly assigned to three treatments in nine replicates, with one rat per litter. Dietary treatments were basal diet (CON), CON + 50 mg/kg 5-ALA (5-ALA50), and CON + 100 mg/kg 5-ALA (5-ALA100). After parturition, ten pups in each litter (a total of 270) were selected for continued feeding by their corresponding mother, and the pregnant rats were fed diets containing 5-ALA (0, 50 and 100 mg/kg diet) until the newborn pups were weaned at 21 days. The results showed that the number of red blood cells (RBCs) in weaned rats in the 5-ALA100 group was significantly higher (p < 0.05) than that in the CON or 5-ALA50 group. The diet with 5-ALA significantly increased (p < 0.05) the hemoglobin (HGB) concentration, hematocrit (HCT) level, serum iron (SI) content, and transferrin saturation (TSAT) level in the blood of weaned rats, as well as the concentration of Hepcidin in the liver and serum of weaned rats and the expression of Hepcidin mRNA in the liver of weaned rats, with the 5-ALA100 group having the highest (p < 0.05) HGB concentration in the weaned rats, and the 5-ALA50 group having the highest (p < 0.05) Hepcidin concentration in serum and in the expression of Hepcidin mRNA in the liver of weaned rats. The other indicators between the 5-ALA groups had no effects. However, the level of total iron binding capacity (TIBC) was significantly decreased (p < 0.05) in the 5-ALA50 group. Moreover, the iron content in the liver of weaned rats fed with 5-ALA showed an upward trend (p = 0.085). In addition, feeding a 5-ALA-supplemented diet could also significantly reduce (p < 0.05) the expression of TfR1 mRNA in the liver of weaning rats (p < 0.05), and the expression of Tfr1 was not affected between 5-ALA groups. In conclusion, dietary supplementation with 5-ALA could improve the blood parameters, increase the concentration of Hepcidin in the liver and serum, and affect the expression of iron-related genes in the liver of weaned rats. Moreover, it is appropriate to add 50 mg/kg 5-ALA to the diet under this condition.

The treatment of neoplastic disease of the brain is still a challenge for modern medicine. Therefore, advanced methodologies are needed that can rationally and successfully contribute to the early diagnosis of primary and metastatic tumors growing within the brain. Photodynamic therapy (PDT) seems to be a valuable method of treatment for precancerous and cancerous lesions including brain tumors. The main advantage of PDT is its high efficiency, minimal invasiveness and no serious side effects, compared with chemotherapy and radiotherapy. This review was conducted through a comprehensive search of articles, scientific information databases and the websites of organizations dealing with cancer treatment. Key points from clinical trials conducted by other researchers are also discussed. The common databases such as PubMed, Google Scholar, EBSCO, Scopus, and Elsevier were used. Articles in the English language of reliable credibility were mainly analyzed. The type of publications considered included clinical and preclinical studies, systematic reviews, and case reports. Based on these collected materials, we see that scientists have already demonstrated the potential of PDT application in the field of brain tumors. Therefore, in this review, the treatment of neoplasm of the Central Nervous System (CNS) and the most common tumor, glioblastoma multiforme (GBM), have been explored. In addition, an overview of the general principles of PDT, as well as the mechanism of action of the therapy as a therapeutic platform for brain tumors, is described. The research was carried out in June 2022.

Bladder cancer is commonly diagnosed by evaluating the tissue morphology through cystoscopy, and tumor resection is used as the primary treatment approach. However, these methods are limited by lesion site specificity and resection margin, and can thereby fail to detect cancer lesions at early stages. Nevertheless, rapid diagnosis without biopsy may be possible through fluorescence sensing. Herein, we describe a minimally invasive imaging system capable of sensing even small tumors through a 1.2 mm diameter flexible fiber bundle microprobe. We demonstrate that this new device can be used for the early diagnosis of bladder cancer in rats. Bladder cancer was induced in rats using the carcinogen N-butyl-N-(4-hydroxybutyl)nitrosamine (BBN), and a togglable filter capable of PpIX fluorescence sensing was installed in the microendoscopic system. Following 5-aminolevulinic acid administration, tissue in the early stages of bladder cancer was successfully identified with fluorescence detection and confirmed with hematoxylin/eosin and ferrochelatase staining. Although the time required for BBN to induce bladder cancer varied between 3 and 4 weeks among the rats, the microendoscopic system allowed the minimally invasive follow-up on cancer development.

Detecting early-stage oral cancer and precancerous lesions are critical to improving patient prognosis and quality of life after treatment. Photodynamic diagnosis using 5-aminolevulinic acid enables the detection of malignant lesions. This study aimed to improve the diagnostic accuracy of photodynamic diagnosis using an objective chromaticity analysis of fluorescence emitted from oral lesions. Sixty-seven patients with clinically suspicious oral cavity lesions underwent photodynamic diagnosis after topical application of 5-aminolevulinic acid solution, followed by imaging and histological evaluation of the lesions. Chromaticity red and green values were measured from the fluorescence images on the lesion, and the red-to-green ratio was calculated. The photodynamic diagnosis allowed for the visualization of oral cancer and high-risk dysplasia as red fluorescence. Compared to low-risk dysplasia and benign lesions, oral cancer and high-risk dysplasia areas had a significantly higher red value and red-to-green ratio. After setting the cutoff value, sensitivity and specificity were 83.3–88.7% and 83.3–83.9%, respectively, when discriminating between oral cancer or high-risk dysplasia and low-risk dysplasia or benign lesions. Photodynamic diagnosis combined with chromaticity analysis may be a valuable diagnostic tool for detecting oral lesions, with a high likelihood of malignant transformation.

Oral squamous-cell and pancreatic carcinomas are aggressive cancers with a poor outcome. Photodynamic therapy (PDT) consists of the use of photosensitizer-induced cell and tissue damage that is activated by exposure to visible light. PDT selectively acts on cancer cells, which have an accumulation of photosensitizer superior to that of the normal surrounding tissues. 5-aminolevulinic acid (5-ALA) induces the production of protoporphyrin IX (PpIX), an endogenous photosensitizer activated in PDT. This study aimed to test the effect of a new gel containing 5% v / v 5-ALA (ALAD-PDT) on human oral CAL-27 and pancreatic CAPAN-2 cancer cell lines. The cell lines were incubated in low concentrations of ALAD-PDT (0.05%, 0.10%, 0.20%, 0.40%, 0.75%, 1.0%) for 4 h or 8 h, and then irradiated for 7 min with 630 nm RED light. The cytotoxic effects of ALAD-PDT were measured using the MTS assay. Apoptosis, cell cycle, and ROS assays were performed using flow cytometry. PpIX accumulation was measured using a spectrofluorometer after 10 min and 24 and 48 h of treatment. The viability was extremely reduced at all concentrations, at 4 h for CAPAN-2 and at 8 h for CAL-27. ALAD-PDT induced marked apoptosis rates in both oral and pancreatic cancer cells. Elevated ROS production and appreciable levels of PpIX were detected in both cell lines. The use of ALA-PDT as a topical or intralesional therapy would permit the use of very low doses to achieve effective results and minimize side effects. ALAD-PDT has the potential to play a significant role in complex oral and pancreatic anticancer therapies.

Aim: The objective of this study was to compare the efficacy of supportive periodontal therapy (i.e., scaling and root planning, SRP) alone versus ALADENT medical device used in association with SRP in the treatment of chronic periodontitis in adult patients. Materials and Methods: A total of 20 patients with a diagnosis of chronic periodontitis (40 localized chronic periodontitis sites) aged between 35 and 55 were selected. None of these patients previously received any surgical or non-surgical periodontal therapy, and they presented radiographic evidence of moderate bone loss. Two non-adjacent sites in different quadrants were identified and observed in each patient, analyzing treatment effectiveness (split-mouth design). Clinical pocket depth, clinical attachment loss, and bleeding on probing were evaluated at time 0 and after 6 months, while microbial analysis (MA) was conducted at baseline and after 15 days. Significant differences were calculated using SPSS program and paired simple statistic t-test. Results: Total bacteria loadings had a statistically significant reduction before and after treatment with SRP (left site) (total average decrease of 27%). The sites treated with SRP plus ALADENT (right) showed a significantly reduced total bacterial loading compared to the untreated sites (right) (total average decrease of 75%). Mean values of CAL/PD and percentages data of BOP, recorded after SRP + ALADENT therapy, showed a higher reduction (CAL = 2.42, PD = 2.87 mm, 90% of sites with no bleeding) than those obtained after SRP treatment (CAL = 4.08 mm, PD = 4.73 mm, 70% of sites with no bleeding). Conclusion: The treatment of moderate and severe chronic periodontitis should include, beside SRP, the use of ALADENT medical device, which has been proved to be a useful adjuvant therapy.

Plant growth, development, and productivity are adversely affected under drought conditions. Previous findings indicated that 5-aminolevulinic acid (ALA) and 24-epibrassinolide (EBL) play an important role in the plant response to adverse environmental conditions. This study demonstrated the role of ALA and EBL on oxidative stress and photosynthetic capacity of drought-stressed ‘Williams’ banana grown under the Egyptian semi-arid conditions. Exogenous application of either ALA or EBL at concentrations of 15, 30, and 45 mg·L−1 significantly restored plant photosynthetic activity and increased productivity under reduced irrigation; this was equivalent to 75% of the plant’s total water requirements. Both compounds significantly reduced drought-induced oxidative damages by increasing antioxidant enzyme activities (superoxide dismutase ‘SOD’, catalase ‘CAT’, and peroxidase ‘POD’) and preserving chloroplast structure. Lipid peroxidation, electrolyte loss and free non-radical H2O2 formation in the chloroplast were noticeably reduced compared to the control, but chlorophyll content and photosynthetic oxygen evolution were increased. Nutrient uptake, auxin and cytokinin levels were also improved with the reduced abscisic acid levels. The results indicated that ALA and EBL could reduce the accumulation of reactive oxygen species and maintain the stability of the chloroplast membrane structure under drought stress. This study suggests that the use of ALA or EBL at 30 mg·L−1 can promote the growth, productivity and fruit quality of drought-stressed banana plants.

This study aimed to establish a method to detect canine urothelial carcinoma cells in urine using 5-aminolevulinic acid (5-ALA) and to evaluate its diagnostic accuracy. Urine samples were collected from 21 dogs diagnosed with urothelial carcinoma and three urothelial carcinoma cell lines were used. Urine samples obtained from seven healthy dogs were used as controls. Cells in the urine sediment, or urothelial carcinoma cell lines, were cultured with 5-ALA and then observed under a fluorescence microscope. Moreover, we examined the relationship between fluorescence intensity and the presence of metastasis as well as tumor invasion into the bladder wall in cases of urothelial carcinoma. Urine-derived cells from urothelial carcinoma and urothelial carcinoma cell lines showed clearer red fluorescence with the addition of 5-ALA compared to that exhibited by the cells from healthy dogs. The sensitivity and specificity of the diagnosis of urothelial carcinoma were 90% and 86%, respectively. Significant associations were found between fluorescence intensity and tumor metastasis and bladder wall invasion. This study showed that 5-ALA can be used to detect urothelial carcinoma cells in dogs with relatively high diagnostic accuracy. Further, the fluorescence intensity of tumor cells caused by 5-ALA correlated with the clinical condition of urothelial carcinoma cases, which suggested that 5-ALA could be used as a prognostic marker for canine urothelial carcinoma.

Photodynamic therapy (PDT) is safe and effective in the treatment of patients with actinic keratosis (AK). The aim of the study was to assess the efficacy, tolerability and cosmetic outcome of topical PDT in the treatment of AKs with three forms of photosensitizers: 5-Aminolevulinic acid hydrochloride (ALA-HCl), 5-Aminolevulinate methyl ester hydrochloride (MAL-HCl) and 5-Aminolevulinate phosphate (ALA-P). The formulations were applied onto selected scalp/face areas. Fluorescence was assessed with a FotoFinder Dermoscope 800 attachment. Skin areas were irradiated with Red Beam Pro+, Model APRO (MedLight GmbH, Herford, Germany). Applied treatments were assessed during the PDT as well as 7 days and 12 weeks after its completion. Ninety-four percent of patients rated obtained cosmetic effect excellent. The efficacy of applied PSs did not differ significantly. However, pain intensity during the PDT procedure was significantly lower in the area treated with ALA-P (5.8 on average) in comparison to the areas treated with ALA-HCl or MAL-HCl (7.0 on average on 0–10 scale). Obtained results show that ALA-P may undergo more selective accumulation than ALA-HCl and MAL-HCl. Our promising results suggest that PDT with the use of ALA-P in AK treatment may be an advantageous alternative to the already used ALA-HCl and MAL-HCl.

The effect of low-dose photodynamic therapy on in vivo wound healing was investigated using optical coherence tomography. This work aims to develop an approach to quantitative assessment of the wound’s state during wound healing including the effect of low-dose photodynamic therapy using topical application of two different photosensitizers, 5-aminolevulinic acid and methylene blue, and two laser doses of 1 J/cm2 and 4 J/cm2. It was concluded that the laser dose of 4 J/cm2 was better compared to 1 J/cm2 and allowed the wound healing process to accelerate.

Alpinia zerumbet is an important medicinal and ornamental plant species. Drought stress is a major concern for sustainable horticulture crop production under changing climate scenarios. Trehalose (Tre) and 5-aminolevulinic acid (ALA) are osmoprotectants that play important roles in mitigating plant stresses. In this study, the effects of foliar application of 25 mM Tre or 10 mg L−1 ALA on biochemical and physiological parameters of A. zerumbet seedlings and their growth were evaluated under well-watered and drought-stressed (65% of field capacity) conditions. Drought caused reductions in physiological parameters and plant growth. These decreases were accompanied by increases in leaf free proline and glycine betaine concentrations and peroxidase activities. Foliar application of Tre or ALA remediated physiological and biochemical parameters and plant growth. Overall, foliar application of ALA or Tre proved to be beneficial for mitigating drought stress in A. zerumbet.

Chemotherapy is cytotoxic to various cancer cells and as well as normal cells. Thus, treatments that demonstrate selective cytotoxicity for cancer cells are desired. The combination of chemotherapy and other cancer therapies can show synergic cytotoxicity, which may be a clue to the nature of the involved cancer cellar-specific damage. We previously reported a phenomenon whereby mitochondrial reactive oxygen species (mitROS) regulate the expression transporters involved in anticancer drug transport and mitROS production is increased by hyperthermia. Moreover, the uptake of 5-aminolevulinic acid (ALA) was enhanced by the increase in mitROS production. In this study, we investigated whether the combination of hyperthermia and ALA can enhance the cytotoxicity of doxorubicin. MitROS production and ALA-derived porphyrin accumulation by hyperthermia (HT) were increased in a murine breast cancer cell line. The expression of solute carrier 15A1 (SLC15A1) upregulated and an ATP-binding cassette subfamily G member 2 (ABCG2) downregulated by HT. Since SLC15A1 is an accumulating transporter for ALA, while ABCG2 is a porphyrin efflux transporter, porphyrin accumulation was enhanced. ABCG2 is also a doxorubicin efflux transporter. Thus, ALA treatment accelerates the intracellular concentration of porphyrin, which acts as a competitive inhibitor of doxorubicin. Indeed, the amount of intracellular doxorubicin was increased by a combination of HT and ALA. The cytotoxicity of doxorubicin was also enhanced. This enhancement was observed in the human breast cancer cell line while it was not seen in normal cells. The combination of HT and ALA treatment can enhance the cancer-specific cytotoxicity of doxorubicin.

Sonodynamic therapy is a bimodal therapeutic approach in which a chemical compound and ultrasound (US) synergistically act to elicit oxidative damage, triggering cancer cell death. Despite encouraging results, mainly for anticancer treatment, sonodynamics is still far from having a clinical application. Therefore, to close the gap between the bench and bedside, more in vivo studies are needed. In this investigation, the combined effect of 5-aminolevulinic acid (Ala), a natural porphyrin precursor, plus exposure to US, was investigated in vivo on a syngeneic breast cancer model. Real-time RT-PCR, Western blotting, and immunohistochemistry assays were performed to evaluate the effect of sonodynamic treatment on the main cancer hallmarks. The sonodynamic-treated group had a significant reduction (p ≤ 0.0001) in tumor size compared to the untreated group, and the Ala- and US-only treated groups, where a strong decrease (p ≤ 0.0001) in Ki67 protein expression was the most relevant feature of sonodynamic-treated cancer tissues. Moreover, oxidative stress was confirmed as the pivotal driver of the anticancer effect through cell cycle arrest, apoptosis, and autophagy; thus, sonodynamics should be explored further for cancer treatment.

Oxidative stress is imparted by a varying range of environmental factors involving heavy metal stress. Thus, the mechanisms of antioxidant resistance may advance a policy to improve metal tolerance. Lead as a toxic heavy metal negatively affects the metabolic activities and growth of medicinal and aromatic plants. This investigation aimed to assess the function of 5-aminolevulinic acid (ALA) in the alleviation of Pb stress in sage plants (Salvia officinalis L.) grown either hydroponically or in pots. Various concentrations of Pb (0, 100, 200, and 400 µM) and different concentrations of ALA (0, 10, and 20 mg L−1) were tested. This investigation showed that Pb altered the physiological parameters. Pb stress differentially reduced germination percentage and protein content compared to control plants. However, lead stress promoted malondialdehyde (MDA) and H2O2 contents in the treated plants. Also, lead stress enhanced the anti-oxidative enzyme activities; ascorbate peroxidase superoxide, dismutase, glutathione peroxidase, and glutathione reductase in Salvia plants. ALA application enhanced the germination percentage and protein content compared to their corresponding controls. Whereas, under ALA application MDA and H2O2 contents, as well as the activities of SOD, APX, GPX, and GR, were lowered. These findings suggest that ALA at the 20 mgL−1 level protects the Salvia plant from Pb stress. Therefore, the results recommend ALA application to alleviate Pb stress.

Extracorporeal photopheresis (ECP), an immunomodulatory therapy for the treatment of chronic graft-versus-host disease (cGvHD), exposes isolated white blood cells to photoactivatable 8-methoxypsoralen (8-MOP) and UVA light to induce the apoptosis of T-cells and, hence, to modulate immune responses. However, 8-MOP-ECP kills diseased and healthy cells with no selectivity and has limited efficacy in many cases. The use of 5-aminolevulinic acid (ALA) and light (ALA-based photodynamic therapy) may be an alternative, as ex vivo investigations show that ALA-ECP kills T-cells from cGvHD patients more selectively and efficiently than those treated with 8-MOP-ECP. The purpose of this phase I-(II) study was to evaluate the safety and tolerability of ALA-ECP in cGvHD patients. The study included 82 treatments in five patients. One patient was discharged due to the progression of the haematological disease. No significant persistent changes in vital signs or laboratory values were detected. In total, 62 adverse events were reported. Two events were severe, 17 were moderate, and 43 were mild symptoms. None of the adverse events evaluated by the internal safety review committee were considered to be likely related to the study medication. The results indicate that ALA-ECP is safe and is mainly tolerated well by cGvHD patients.

Photodynamic therapy (PDT) has been investigated as an effective, non-invasive, and alternative tumor-ablative therapy that uses photosensitizers (PSs) and safe irradiation light in the presence of oxygen to generate reactive oxygen species (ROS) to kill malignant cancer cells. However, the off-target activation of the PSs can hinder effective PDT. Therefore, an advanced drug delivery system is required to selectively deliver the PS to the therapeutic region only and reduce off-target side effects in cancer treatment. The integration of laser-initiated PDT with nanotechnology has provided new opportunities in cancer therapy. In this study, plasmonic bimetallic nanoparticles (NPs) were prepared for the targeted PDT (TPDT) of in vitro cultured MCF-7 breast cancer cells. The NPs were functionalized with PEG through Au–thiol linkage to enhance their biocompatibility and subsequently attached to the PS precursor 5-aminolevulinic acid via electrostatic interactions. In order to enhance specific targeting, anti-HER-2 antibodies (Ab) were decorated onto the surface of the nanoconjugate (NC) to fabricate a 5-ALA/Au–Ag-PEG-Ab NC. In vitro studies showed that the synthesized NC can enter MCF-7 cells and localize in the cytoplasm to metabolize 5-ALA to protoporphyrin IX (PpIX). Upon light irradiation, PpIX can efficiently produce ROS for the PDT treatment of MCF-7. Cellular viability studies showed a decrease from 49.8% ± 5.6 ** to 13.8% ± 2.0 *** for free 5-ALA versus the NC, respectively, under equivalent concentrations of the PS (0.5 mM, IC50). These results suggest that the active targeted NC platform has an improved PDT effect on MCF-7 breast cancer cells.

Heme enzyme dysfunction causes a group of diseases called porphyrias. Particularly, a decrease in porphobilinogen deaminase, involved in the third step of heme biosynthesis, leads to acute intermittent porphyria (AIP). Considering our previous works demonstrating the multiplicity of brain metabolisms affected by porphyrinogenic agents, this study aimed to elucidate whether they cause any alteration on the mitochondrial respiratory chain. The activities of respiratory chain complexes (I to IV) were measured in encephalon mitochondria of CF1 male mice receiving volatile anesthetics: isoflurane (2 mL/kg) and sevoflurane (1.5 mL/kg), ethanol (30%), allylisopropylacetamide (AIA) (350 mg/kg), and barbital (167 mg/kg). Moreover, they were compared versus animals with pathological levels of 5-aminolevulinic acid (ALA, 40 mg/kg). Complex I-III activity was induced by isoflurane and decreased by AIA, ethanol, and ALA. Complex II-III activity was increased by sevoflurane and decreased by isoflurane and AIA. Complex II activity was increased by sevoflurane and barbital and decreased by AIA, ethanol, and ALA. Complex IV activity was increased by barbital and ALA and decreased by sevoflurane. The damage to the respiratory chain by ALA could be reflecting the pathophysiological condition of patients with AIP. Better understanding the broad effect of porphyrinogenic drugs and the mechanisms acting on the onset of AIP is vital in translational medicine.

It is well known that trees grown on roadsides suffer from stressful environments, including poor soils, bad weather, and harmful gases from automobile exhaust. Improving the adaptability of roadside trees to adverse environments is important for urban management. An experiment was carried out with six-year-old Buxus megistophylla Levl. hedgerows, where 20 mg/L 5-aminolevulinic acids (5-ALA) solution was sprayed on the blade surface at the end of April. Three months later, plant morphology, chlorophyll fast fluorescence characteristics, antioxidant enzyme activities and the mineral element content were investigated. The results showed that leaf size and thickness were significantly greater with 5-ALA treatment, and the leaf color was also greener than those of the control. 5-ALA treatment significantly promoted the electron transfer activity of the PSII reaction center on the donor side, the reaction center itself and the receptor side. It reduced energy dissipation through the heat with increased photochemical quantum yields. The activities of antioxidant enzymes, including superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD) in leaves and roots, were stimulated by 5-ALA treatment. The content of soluble sugars and free proline in leaves was significantly increased by 5-ALA treatment, as were the absorption and accumulation of several kinds of mineral nutrient elements, such as nitrogen, phosphate, calcium, magnesium, iron, copper and boron. Additionally, 5-ALA application significantly increased the content of cadmium, mercury, chromium and lead in the roots but decreased them in the leaves. This implies that 5-ALA may induce a mechanism in B. megistophylla in which toxic elements were intercepted in roots to avoid accumulation in leaves, which ensured healthy growth of the aboveground tissues. 5-ALA may regulate the absorption and utilization of mineral nutrient elements in soil with the interception of toxic heavy metal elements in roots, promote leaf photosynthetic performance, induce the accumulation of soluble sugars and free proline, and improve the antioxidant enzyme systems for plants to adapt to the stressful environment of urban roads. These results provide a basis for 5-ALA applications alongside city roads.

Chronic and infected leg ulcers (LUs) are painful, debilitating, resistant to antibiotics, and immensely reduce a patient’s quality of life. The purpose of our study was to demonstrate the efficacy of photodynamic therapy (PDT) for the treatment of infected chronic LUs. Patients were randomized into two experimental groups: the first group received 5-aminolevulinic acid photodynamic therapy (ALA-PDT) (10 patients), and the second group of 10 patients received local octenidine dihydrochloride (Octenilin gel) exposed to a placebo light source with an inserted filter that mimiced red light. In the PDT group, we used 20% ALA topically applied for 4 hrs and irradiation from a Diomed laser source with a wavelength of 630 nm at a fluency of 80 J/cm2. ALA-PDT was performed 10 times during a 14-day hospitalization in 10 patients of both sexes aged 40–85 years with chronic leg ulcers. Treatments were carried out at 3-week intervals for 3–5 cycles. At 8-month follow-up with the PDT group, complete remission (CR) was obtained in four patients (40%), partial response (>50% reduction in ulcer diameter) in four patients (40%), and no response in two patients (20%) who additionally developed deterioration of the local condition with swelling, erythema, and inflammation. To assess the degree of pain during the trials, we used a numeric rating scale (NRS). From the preliminary results obtained, we concluded that PDT can be used to treat leg ulcers as a minimally invasive and effective method with no serious side effects, although further studies on a larger group of patients with LUs are warranted.

5-Aminolevulinic acid (5-ALA) is an amino acid derivative and a precursor of protoporphyrin IX (PpIX). The photophysical feature of PpIX is clinically used in photodynamic diagnosis (PDD) and photodynamic therapy (PDT). These clinical applications are potentially based on in vitro cell culture experiments. Thus, conducting a systematic review and meta-analysis of in vitro 5-ALA PDT experiments is meaningful and may provide opportunities to consider future perspectives in this field. We conducted a systematic literature search in PubMed to summarize the in vitro 5-ALA PDT experiments and calculated the effectiveness of 5-ALA PDT for several cancer cell types. In total, 412 articles were identified, and 77 were extracted based on our inclusion criteria. The calculated effectiveness of 5-ALA PDT was statistically analyzed, which revealed a tendency of cancer-classification-dependent sensitivity to 5-ALA PDT, and stomach cancer was significantly more sensitive to 5-ALA PDT compared with cancers of different origins. Based on our analysis, we suggest a standardized in vitro experimental protocol for 5-ALA PDT.

Clinical evidence regarding risk reduction of repeated bladder recurrence after initial photodynamic diagnosis (PDD)-assisted transurethral resection of bladder tumor (TURBT) is still limited in patients with non-muscle-invasive bladder cancer (NMIBC). We analyzed patients with primary NMIBC undergoing TURBT without any adjuvant treatment to compare the risk of cumulative recurrence between the conventional white-light (WL)-TURBT and PDD-TURBT. Out of 430 patients diagnosed with primary NMIBC from 2010 to 2019, 40 undergoing WL-TURBT and 60 undergoing PDD-TURBT were eligible. Multivariate Cox regression analysis for time to the first recurrence demonstrated that PDD assistance was an independent prognostic factor with better outcome (p = 0.038, hazard ratio = 0.39, and 95% confidence interval 0.16–0.95). While no patient experienced more than one recurrence within 1000 postoperative days in the PDD-TURBT group, five out of 40 patients treated by WL-TURBT experienced repeated recurrence. The comparison of cumulative incidence per 10,000 person-days between the two groups revealed that PDD assistance decreased by 6.6 recurrences per 10,000 person-days (exact p = 0.011; incidence rate ratio 0.37, Clopper–Pearson confidence interval 0.15–0.82). This is the first study addressing PDD assistance-induced risk reduction of repeated bladder recurrence using the person-time method. Our findings could support clinical decision making, especially on adjuvant therapy after TURBT.

Background: A very important aspect in the treatment of high-grade glioma is gross total resection to reduce the risk of tumor recurrence. One of the methods to facilitate this task is intraoperative fluorescence navigation. The aim of the study was to compare the dyes used in this technique fluorescent intraoperative navigation in terms of the mechanism of action and influence on the treatment of patients. Methods: The review was carried out on the basis of articles found in PubMed, Google Scholar, and BMC search engines, as well as those identified by searched bibliographies and suggested by experts during the preparation of the article. The database analysis was performed for the following phrases: “glioma”, “glioblastoma”, “ALA”, “5ALA”, “5-ALA”, “aminolevulinic acid”, ”levulinic acid”, “fluorescein”, “ICG”, “indocyanine green”, and “fluorescence navigation”. Results: After analyzing 913 citations identified on the basis of the search criteria, we included 36 studies in the review. On the basis of the analyzed articles, we found that 5-aminolevulinic acid and fluorescein are highly effective in improving the percentage of gross total resection achieved in high-grade glioma surgery. At the same time, the limitations resulting from the use of these methods are marked—higher costs of the procedure and the need to have neurosurgical microscope in combination with a special light filter in the case of 5-aminolevulinic acid (5-ALA), and low specificity for neoplastic cells and the dependence on the degree of damage to the blood–brain barrier in the intensity of fluorescence in the case of fluorescein. The use of indocyanine green in the visualization of glioma cells is relatively unknown, but some researchers have suggested its utility and the benefits of using it simultaneously with other dyes. Conclusion: The use of intraoperative fluorescence navigation with the use of 5-aminolevulinic acid and fluorescein allows the range of high-grade glioma resection to be increased.

In cancer nanomedicine, numerous studies have been conducted on the surface modification and transport capacity of nanoparticles (NPs); however, biological barriers, such as enzymatic degradation or non-specific delivery during circulation, remain to be cleared. Herein, we developed pH-sensitive NPs that degrade in an acidic environment and release 5-aminolevulinic acid (5ALA) to the target site. NPs were prepared by conjugating alginate with folic acid, followed by encapsulation of 5ALA through a water-in-oil (W/O) emulsion method. The alginate-conjugated folic acid nanoparticles (AF NPs) were homogeneous in size, stable for a long time in aqueous suspension without aggregation, and non-toxic. AF NPs were small enough to efficiently infiltrate tumors (

Prognosis for diffuse intrinsic pontine glioma (DIPG) and generally for diffuse midline gliomas (DMG) has only marginally improved over the last ~40 years despite dozens of chemotherapy and other therapeutic trials. The prognosis remains invariably fatal. We present here the rationale for a planned study of adding 5-aminolevulinic acid (5-ALA) to the current irradiation of DIPG or DMG: the 5aai regimen. In a series of recent papers, oral 5-ALA was shown to enhance standard therapeutic ionizing irradiation. 5-ALA is currently used in glioblastoma surgery to enable demarcation of overt tumor margins by virtue of selective uptake of 5-ALA by neoplastic cells and selective conversion to protoporphyrin IX (PpIX), which fluoresces after excitation by 410 nm (blue) light. 5-ALA is also useful in treating glioblastomas by virtue of PpIX’s transfer of energy to O2 molecules, producing a singlet oxygen that in turn oxidizes intracellular DNA, lipids, and proteins, resulting in selective malignant cell cytotoxicity. This is called photodynamic treatment (PDT). Shallow penetration of light required for PpIX excitation and resultant energy transfer to O2 and cytotoxicity results in the inaccessibility of central structures like the pons or thalamus to sufficient light. The recent demonstration that keV and MeV photons can also excite PpIX and generate singlet O2 allows for reconsideration of 5-ALA PDT for treating DMG and DIPG. 5-ALA has an eminently benign side effect profile in adults and children. A pilot study in DIPG/DMG of slow uptitration of 5-ALA prior to each standard irradiation session—the 5aai regimen—is warranted.

Heme, like chlorophyll, is a primordial molecule and is one of the fundamental pigments of life. Disorders of normal heme synthesis may cause human diseases, including certain anemias (X-linked sideroblastic anemias) and porphyrias. Porphyrias are classified as hepatic and erythropoietic porphyrias based on the organ system in which heme precursors (5-aminolevulinic acid (ALA), porphobilinogen and porphyrins) are chiefly overproduced. The hepatic porphyrias are further subdivided into acute porphyrias and chronic hepatic porphyrias. The acute porphyrias include acute intermittent, hereditary copro-, variegate and ALA dehydratase deficiency porphyria. Chronic hepatic porphyrias include porphyria cutanea tarda and hepatoerythropoietic porphyria. The erythropoietic porphyrias include congenital erythropoietic porphyria (Gűnther’s disease) and erythropoietic protoporphyria. In this review, we summarize the key features of normal heme synthesis and its differing regulation in liver versus bone marrow. In both organs, principal regulation is exerted at the level of the first and rate-controlling enzyme, but by different molecules (heme in the liver and iron in the bone marrow). We also describe salient clinical, laboratory and genetic features of the eight types of porphyria.

The purpose of this article is to review the diagnostic possibilities of 5-aminolevulinic acid (5-ALA)-based fluorescence diagnosis of preinvasive cervical changes. Reviewed papers were selected from the PubMed database with keywords combining the terms individual cervical neoplasia and fluorescence diagnostics. The regular colposcopy procedure lacks specificity; therefore, new methods are continually sought for superior diagnosis of cervical pathology. 5-ALA-based fluorescence diagnostics is under investigation as an up-to-date diagnostic technique for cervical intraepithelial neoplasia (CIN). This method is grounded on the topical or systemic application of 5-ALA, which induces excess production of the endogenous photosensitizer protoporphyrin IX (PpIX) in tissues where carcinogenesis has begun. The conversion of PpIX to the heme is less efficient in tumors; therefore, higher amounts of PpIX tend to accumulate in premalignant and malignant tissues. Illumination with light of the appropriate wavelength initiates excitation of PpIX fluorescence, which in turn helps to localize PpIX-rich areas and identify potentially malignant tissues. A number of investigations suggest that because of its high selectivity for tumors and low toxicity to healthy tissues, 5-ALA-based diagnosis seems a promising tool for the noninvasive identification of cervical intraepithelial neoplasia.

Photodynamic therapy (PDT) and photodiagnosis (PD) are essential approaches in the field of biophotonics. Ideally, both modalities require the selective sensitization of the targeted disease in order to avoid undesired phenomena such as the destruction of healthy tissue, skin photosensitization, or mistaken diagnosis. To a large extent, the occurrence of these incidents can be attributed to “background” accumulation in non-target tissue. Therefore, an ideal photoactive compound should be optically silent in the absence of disease, but bright in its presence. Such requirements can be fulfilled using innovative prodrug strategies targeting disease-associated alterations. Here we will summarize the elaboration, characterization, and evaluation of approaches using polymeric photosensitizer prodrugs, nanoparticles, micelles, and porphysomes. Finally, we will discuss the use of 5-aminolevulinc acid and its derivatives that are selectively transformed in neoplastic cells into photoactive protoporphyrin IX.