Search Results (24)

Background: Chronic and acute wounds are often colonized by polymicrobial biofilms, delaying healing and complicating treatment. Rapid, non-invasive detection of pathogenic bacteria is therefore crucial for timely and targeted therapy. This study investigated porphyrin-producing bacterial species using the handheld cureVision imaging system. Methods: In this study, 20 clinically relevant, porphyrin-producing bacterial species were cultured on δ-aminolevulinic acid (ALA)-supplemented agar and analyzed using the handheld cureVision imaging system under 405 nm excitation. Both Red-Green-Blue (RGB) and fluorescence images were acquired under ambient daylight conditions, and fluorescence signals were quantified by grayscale intensity analysis. Results: All tested species exhibited measurable red porphyrin-associated fluorescence, with the highest intensities observed in Klebsiella pneumoniae, Klebsiella oxytoca, Veillonella parvula, and Alcaligenes faecalis. A standardized detectability threshold of 0.25, derived from negative controls, enabled semi-quantitative comparison across species. Statistical analysis confirmed that the fluorescence intensities of all bacterial samples were significantly elevated compared to the control (Wilcoxon signed-rank test and sign test, both p < 0.001; median intensity = 0.835, IQR: 0.63–0.975). Conclusions: These results demonstrate that the cureVision system enables robust and reliable detection of porphyrin-producing wound bacteria, supporting its potential as a rapid, non-invasive diagnostic method for assessing wound colonization and guiding targeted clinical interventions. Full article

As natural bioactive compounds, plant pigments play crucial roles not only in plant phenotype, growth, development, and adaptation to stress but also hold unique value in biotechnology, healthcare, and industrial applications. There is growing interest in the biosynthesis and acquisition of plant pigments. Thus, this paper explores emerging extraction methods of natural pigments and elucidates the biosynthesis pathways of four key plant pigments, chlorophylls, carotenoids, anthocyanins, and betalaine in vivo and in vitro. We comprehensively discuss the application of solvent, supercritical fluid [extraction], ultrasonic, and microwave-assisted extraction techniques, as well as introducing key enzymes, precursors, and synthetic pathways involved in pigment synthesis. δ-Aminolevulinic acid represents a pivotal initiating enzyme for chlorophyll synthesis, whereas isopentenylpyrophosphate, (IPP) and dimethylallyl pyrophosphate, (DMAPP) are closely associated with carotenoid biosynthesis. Phenylalanine and tyrosine are critical substances for anthocyanin and betalaine synthesis, respectively. Hence, crucial genes such as chlI, crtB, PGT8, CYP76AD1, and BvDODA can be employed for heterologous biosynthesis in vitro to meet the demand for increased plant pigment amount. As a pivotal determinant of plant coloration, an in-depth exploration into the high-quality acquisition of plant pigments can provide a basis for developing superior pigments and offer new insights into increasing pigment yield. Full article

Feline leukemia virus C receptor 1a (FLVCR1a), initially identified as a retroviral receptor and localized on the plasma membrane, has emerged as a crucial regulator of heme homeostasis. Functioning as a positive regulator of δ-aminolevulinic acid synthase 1 (ALAS1), the rate-limiting enzyme in the heme biosynthetic pathway, FLVCR1a influences TCA cycle cataplerosis, thus impacting TCA flux and interconnected metabolic pathways. This study reveals an unexplored link between FLVCR1a, heme synthesis, and cholesterol production in endothelial cells. Using cellular models with manipulated FLVCR1a expression and inducible endothelial-specific Flvcr1a-null mice, we demonstrate that FLVCR1a-mediated control of heme synthesis regulates citrate availability for cholesterol synthesis, thereby influencing cellular cholesterol levels. Moreover, alterations in FLVCR1a expression affect membrane cholesterol content and fluidity, supporting a role for FLVCR1a in the intricate regulation of processes crucial for vascular development and endothelial function. Our results underscore FLVCR1a as a positive regulator of heme synthesis, emphasizing its integration with metabolic pathways involved in cellular energy metabolism. Furthermore, this study suggests that the dysregulation of heme metabolism may have implications for modulating lipid metabolism. We discuss these findings in the context of FLVCR1a’s potential heme-independent function as a choline importer, introducing additional complexity to the interplay between heme and lipid metabolism. Full article

Erythropoietic protoporphyria (EPP) is a genetic disorder stemming from reduced ferrochelatase expression, the final enzyme in the pathway of heme biosynthesis. A closely related condition, X-linked protoporphyria (XLP), bears similar clinical features although it arises from the heightened activity of δ-aminolevulinic acid synthase 2 (ALAS2), the first and normally rate-controlling enzyme in heme biosynthesis in developing red blood cells. Both of these abnormalities result in the buildup of protoporphyrin IX, leading to excruciating light sensitivity and, in a minority of cases, potentially fatal liver complications. Traditionally, managing EPP and XLP involved sun avoidance. However, the emergence of innovative therapies, such as dersimelagon, is reshaping the therapeutic landscape for these conditions. In this review, we summarize salient features of the properties of dersimelagon, shedding light on its potential role in advancing our understanding of treatment options for EPP and XLP. Full article

Redox regulation of plastid gene expression and different metabolic pathways promotes many activities of redox-sensitive proteins. We address the question of how the plastid redox state and the contributing reducing enzymes control the enzymes of tetrapyrrole biosynthesis (TBS). In higher plants, this metabolic pathway serves to produce chlorophyll and heme, among other essential end products. Because of the strictly light-dependent synthesis of chlorophyll, tight control of TBS requires a diurnal balanced supply of the precursor 5-aminolevulinic acid (ALA) to prevent the accumulation of photoreactive metabolic intermediates in darkness. We report on some TBS enzymes that accumulate in a light intensity-dependent manner, and their contents decrease under oxidizing conditions of darkness, low light conditions, or in the absence of NADPH-dependent thioredoxin reductase (NTRC) and thioredoxin f1 (TRX-f1). Analysis of single and double trxf1 and ntrc mutants revealed a decreased content of the early TBS enzymes glutamyl-tRNA reductase (GluTR) and 5-aminolevulinic acid dehydratase (ALAD) instead of an exclusive decrease in enzyme activity. This effect was dependent on light conditions and strongly attenuated after transfer to high light intensities. Thus, it is suggested that a deficiency of plastid-localized thiol-redox transmitters leads to enhanced degradation of TBS enzymes rather than being directly caused by lower catalytic activity. The effects of the proteolytic activity of the Clp protease on TBS enzymes were studied by using Clp subunit-deficient mutants. The simultaneous lack of TRX and Clp activities in double mutants confirms the Clp-induced degradation of some TBS proteins in the absence of reductive activity of TRXs. In addition, we verified previous observations that decreased chlorophyll and heme levels in ntrc could be reverted to WT levels in the ntrc/Δ2cp triple mutant. The decreased synthesis of 5-aminolevulinic acid and porphobilinogen in ntrc was completely restored in ntrc/Δ2cp and correlated with WT-like levels of GluTR, ALAD, and other TBS proteins. Full article

The present study aimed to identify the abilities of the δ-aminolevulinic acid (ALA) producing purple non-sulfur bacteria (PNSB), Luteovulum sphaeroides W22 and W47, to reduce the Na⁺ concentration, and to ameliorate the soil fertility, nutrients uptake, growth and yield of rice on the salt-contaminated soil. A two-factor experiment was conducted following a completely randomized block design. The factors were the frequency of applying saline irrigation (zero, one, two, three and four times) and the ALA-producing PNSB supplementation (applying only W22, only W47 and mixed W22 + W47). The results revealed that supplying the PNSB mixture not only reduced the proline content but also increased the plant height, number of panicles per pot, percentage of filled seeds, contents of NH₄⁺, PO₄³⁻, total N, P uptake and grain yield. The mixed PNSB application also reduced the Na content and the total Na uptake in plants. L. sphaeroides W22 and W47 decreased the proline content by 31.3% and increased the grain yield by 27.2% in the condition of applying 5‰ saline irrigation four times. Full article

High elimination rates and concerns for horse welfare are important issues in endurance riding. Improved understanding of the causes of elimination could increase completion rates in this sport. We have identified pre-ride laboratory risk factors that enable an assessment of potential elimination before the ride. A longitudinal cohort study was performed among 49 healthy horses competing in the 160 km endurance ride at the 2016 World Championship of Endurance Riding in Samorin/Slovakia. Blood samples were taken before the event. For statistical evaluation, horses were categorized into three groups: finishers, lame horses, and metabolically eliminated horses. Risk factors were calculated for each group using multinominal logistic regression. δ-Aminolevulinic-dehydratase (ALAD), thiobarbituric acid reactive substances (TBARSs), iron, and serum amyloid A (SAA) were measured and did not show an impact on the race outcome, but elevated pre-ride superoxide dismutase (SOD) was shown to have an effect on lameness elimination (p = 0.011). It might serve as an indicator for withdrawing horses at risk of later elimination before endurance rides, ultimately resulting in lower elimination rates and an increase in overall horse welfare. Full article

Peaches/nectarines (Prunus persica L.) are widely cultivated worldwide. As with other species, the sugar content is the most important trait for fruit quality, especially for precocious cultivars. Most fruits need to improve their sugar content in order to be more profitable under fierce market competition. 5-Aminolevulinic acid (ALA), a naturally occurring δ-amino acid, has been shown to improve leaf photosynthesis and fruit quality, especially sugar content. However, the mechanisms are not clear. The objective of this study is to determine the effects of exogenous ALA on leaf photosynthesis, assimilate transport, and sugar accumulation during fruit development. We used the field-cultivated precocious nectarine ‘Zhongyoutao 4’ and potted cultivated peach ‘Zhongai 33’ as materials, whereas in the second experiment, we used ¹⁴C radiolabeling to trace ¹⁴C fixation in leaves, transport in branches, and distribution in different organs. The results showed that ALA significantly enhanced the photosynthetic gas exchange capacity, and the effects were maintained for at least one month. The results of the ¹⁴C radiolabel experiment showed that ALA enhanced ¹⁴C fixation in leaves, promoted the transport to fruits, and reduced the allocation rate of young leaves. This suggests that ALA enlarges “source” volume and strengthens “sink” competition; therefore, assimilate translocation to fruits is promoted. It was observed that sucrose contributed the main saccharide for peach fruit quality at maturity, which might not be converted from glucose or fructose but from starch degradation. ALA improved starch accumulation in the young fruits as well as degradation during maturity. The RT-qPCR showed that the expression of most genes involved in sugar metabolism did not correlate or even negatively correlate with fruit sucrose content. However, the expressions of SWEET1/6/7/8/15/16/17 were highly correlated with the sucrose content, and exogenous ALA treatment up-regulated the gene expression at fruit maturity, suggesting they might play an important role in fruit sugar accumulation. These results provide important theoretical support for ALA application in fruit quality improvement, as well as a regulatory mechanism study on sugar accumulation in fruits. Full article

To redirect carbon flux from the γ-aminobutyric acid (GABA) shunt to the δ-aminolevulinic acid (ALA) biosynthetic pathway, we disrupted the GABA shunt route of the model cyanobacterium Synechocystis sp. PCC 6803 by inactivating Gdc, the gene-encoding glutamate decarboxylase. The generated ΔGdc strain exhibited lower intracellular GABA and higher ALA levels than the wild-type (WT) one. The ΔGdc strain’s ALA levels were ~2.8 times higher than those of the WT one when grown with levulinic acid (LA), a competitive inhibitor of porphobilinogen synthase. Abiotic stress conditions including salinity induced by 10 mM NaCl and cold at 4 °C increased the ALA levels in ΔGdc up to ~2.5 and 5 ng g⁻¹ cell DW, respectively. The highest ALA production in the ΔGdc cyanobacteria grown in BG11 medium was triggered by glucose induction, followed by glutamate supplementation with 60 mM of LA, thereby resulting in ~360 ng g⁻¹ cell DW of ALA, that is >300-fold higher ALA accumulation than that observed in ΔGdc cyanobacteria grown in normal medium. Increased levels of the gdhA (involved in the interconversion of α-ketoglutarate to glutamate) and the hemA (a major regulatory target of the ALA biosynthetic pathway) transcripts occurred in ΔGdc cyanobacteria grown under modified growth conditions. Our study provides critical insight into the facilitation of ALA production in cyanobacteria. Full article

Porphobilinogen deaminase (PBGD) haploinsufficiency (acute intermittent porphyria, AIP) is characterized by neurovisceral attacks associated with high production, accumulation and urinary excretion of heme precursors, δ-aminolevulinic acid (ALA) and porphobilinogen (PBG). The estimated clinical penetrance for AIP is extremely low (<1%), therefore it is likely that other factors may play an important role in the predisposition to developing attacks. Fasting is a known triggering factor. Given the increased prevalence of insulin resistance in patients and the large urinary loss of succinyl-CoA to produce ALA and PBG, we explore the impact of reduced availability of energy metabolites in the severity of AIP pathophysiology. Classic studies found clinical improvement in patients affected by AIP associated with the administration of glucose and concomitant insulin secretion, or after hyperinsulinemia associated with diabetes. Molecular studies have confirmed that glucose and insulin administration induces a repressive effect on hepatic ALA Synthase, the first and regulatory step of the heme pathway. More recently, the insulin-mimicking α-lipoic acid has been shown to improve glucose metabolism and mitochondrial dysfunction in a hepatocyte cell line transfected with interfering RNA targeting PBGD. In AIP mice, preventive treatment with an experimental fusion protein of insulin and apolipoprotein A-I improved the disease by promoting fat mobilization in adipose tissue, increasing the metabolite bioavailability for the TCA cycle and inducing mitochondrial biogenesis in the liver. In this review, we analyze the possible mechanisms underlying abnormal hepatocellular carbohydrate homeostasis in AIP. Full article

Salt injury, as an abiotic stress factor, seriously affects the quality and yield of crops. This study was conducted by analyzing alternative splicing in the control (CK), δ-aminolevulinic acid (ALA), NaCl, and ALA + NaCl treatments of wild jujube (Zizyphus spinosus (Bunge)Hu) using RNA-seq. It was found that the unique differential alternative splicing is closely related to the alleviation of salt stress and the analyzed intermediates of chlorophyll synthesis and chlorophyll content in the leaves. The results showed that the content and synthesis of chlorophyll were reduced and disrupted in wild jujube leaves under NaCl stress, and the exogenous spraying of ALA could alleviate the effect of NaCl stress on the content of chlorophyll. RNA-seq indicated that the alternative splicing of genes was dominated by exon skipping in all the experimental treatments. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses revealed that the CK and ALA + NaCl comparison groups were primarily enriched in porphyrin and chlorophyll metabolism, chloroplast, and energy metabolism pathways. It is hypothesized that ALA treatment can alleviate the effects of salt stress on chlorophyll by altering the alternative splicing of genes related to photosynthesis, chlorophyll metabolism, and energy metabolism in wild jujube. In addition, the verification of alternative splicing genes related to chlorophyll metabolism in wild jujube revealed that treatment with ALA significantly upregulated the expression of HEMH, UROIII, and ChlD genes in the leaves under salt stress and increased the content of the chlorophyll biosynthetic intermediates endogenous ALA, porphobilinogen, protoporphyrin IX, Mg-protoporphyrin IX, and protochlorophyllide, which served to alleviate the effects of NaCl stress on chlorophyll in wild jujube. Full article

This work aimed to study the toxic implications of zinc oxide nanoparticles (ZnO NPs) on the physio-biochemical responses of spring barley (Hordeum sativum L.). The experiments were designed in a hydroponic system, and H. sativum was treated with two concentrations of ZnO NPs, namely 300 and 2000 mg/L. The findings demonstrated that ZnO NPs prevent the growth of H. sativum through the modulation of the degree of oxidative stress and the metabolism of antioxidant enzymes. The results showed increased malondialdehyde (MDA) by 1.17- and 1.69-fold, proline by 1.03- and 1.09-fold, and catalase (CAT) by 1.4- and 1.6-fold in shoots for ZnO NPs at 300 and 2000 mg/L, respectively. The activity of superoxide dismutase (SOD) increased by 2 and 3.3 times, ascorbate peroxidase (APOX) by 1.2 and 1.3 times, glutathione-s-transferase (GST) by 1.2 and 2.5 times, and glutathione reductase (GR) by 1.8 and 1.3 times in roots at 300 and 2000 mg/L, respectively. However, the level of δ-aminolevulinic acid (ALA) decreased by 1.4 and 1.3 times in roots and by 1.1 times in both treatments (nano-300 and nano-2000), respectively, indicating changes in the chlorophyll metabolic pathway. The outcomes can be utilized to create a plan of action for plants to withstand the stress brought on by the presence of NPs. Full article

In a course of metabolic experiments, we determined that the addition of δ-aminolevulinic acid (5-ALA) to a panel of glioblastoma multiforme (GBM) cells caused a steep reduction in their glycolytic activity. This reduction was accompanied by a decrease in adenosine triphosphate (ATP) production from glycolysis. These results suggested that 5-ALA is an inhibitor of glycolysis; due to the structural similarity of 5-ALA to the established lactate dehydrogenase (LDH) inhibitors oxamate (OXM) and tartronate (TART), we initially investigated LDH inhibition by 5-ALA in silico. The modelling revealed that 5-ALA could indeed be a competitive inhibitor of LDH but not a substrate. These theoretical findings were corroborated by enzymatic and cell lysate assays in which 5-ALA was found to confer a potent LDH inhibition comparable to that of OXM and TART. We subsequently evaluated the effect of 5-ALA-induced glycolysis inhibition on the viability of GBM cells with diverse metabolic phenotypes. In the Warburg-type cell lines Ln18 and U87, incubation with 5-ALA elicited profound and irreversible cell death (90–98%) at 10 mM after merely 24 h. In T98G, however, which exhibited both high respiratory and glycolytic rates, LD95 was achieved after 72 h of incubation with 20 mM 5-ALA. We additionally examined the production of the 5-ALA photosensitive metadrug protoporphyrin IX (PpIX), with and without prior LDH inhibition by TART. These studies revealed that ~20% of the 5-ALA taken up by the cells was engaged in LDH inhibition. We subsequently performed 5-ALA photodynamic therapy (PDT) on Ln18 GBM cells, again with and without prior LDH inhibition with TART, and found a PDT outcome enhancement of ~15% upon LDH pre-inhibition. We expect our findings to have a profound impact on contemporary oncology, particularly for the treatment of otherwise incurable brain cancers such as GBM, where the specific accumulation of 5-ALA is very high compared to the surrounding normal tissue. Full article

Exposure to lead in environmental and occupational settings continues to be a serious public health problem. At environmentally relevant doses, two mechanisms may underlie lead exposition-induced genotoxicity, disruption of the redox balance and an interference with DNA repair systems. The aim of the study was to evaluate the ability of lead exposition to induce impaired function of Ape1 and its impact on DNA repair capacity of workers chronically exposed to lead in a battery recycling plant. Our study included 53 participants, 37 lead exposed workers and 16 non-lead exposed workers. Lead intoxication was characterized by high blood lead concentration, high lipid peroxidation and low activity of delta-aminolevulinic acid dehydratase (δ-ALAD). Relevantly, we found a loss of DNA repair capacity related with down-regulation of a set of specific DNA repair genes, showing specifically, for the first time, the role of Ape1 down regulation at transcriptional and protein levels in workers exposed to lead. Additionally, using a functional assay we found an impaired function of Ape1 that correlates with high blood lead concentration and lipid peroxidation. Taken together, these data suggest that occupational exposure to lead could decrease DNA repair capacity, inhibiting the function of Ape1, as well other repair genes through the regulation of the ZF-transcription factor, promoting the genomic instability. Full article

Background Acute hepatic porphyrias (AHPs) are a group of rare diseases caused by dysfunctions in the pathway of heme biosynthesis. Although acute neurovisceral attacks are the most dramatic manifestations, patients are at risk of developing long-term complications, several of which are of a vascular nature. The accumulation of non-porphyrin heme precursors is deemed to cause most clinical symptoms. Aim We measured the serum levels of endothelin-1 (ET-1) and nitric oxide (NO) to assess the presence of endothelial dysfunction (ED) in patients with AHPs. Forty-six patients were classified, according to their clinical phenotype, as symptomatic (AP-SP), asymptomatic with biochemical alterations (AP-BA), and asymptomatic without biochemical alterations (AP-AC). Results Even excluding those under hemin treatment, AP-SP patients had the lowest NO and highest ET-1 levels, whereas no significant differences were found between AP-BA and AP-AC patients. AP-SP patients had significantly more often abnormal levels of ED markers. Patients with the highest heme precursor urinary levels had the greatest alterations in ED markers, although no significant correlation was detected. Conclusions ED is more closely related to the clinical phenotype of AHPs than to their classical biochemical alterations. Some still undefined disease modifiers may possibly determine the clinical picture of AHPs through an effect on endothelial functions. Full article