This study explored the self-removal efficiency and mechanisms of a photodynamically mediated 2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPO)-oxidized chitin grafted with 5-aminolevulinic acid (ALA) film against Vibrio parahaemolyticus biofilms via an endogenous photosensitization pathway. Results showed that the grafted ALA induced the production of endogenous protoporphyrin IX (PpIX), which in situ generated much reactive oxygen species (ROS) under blue LED irradiation. The ROS damaged the structural integrity and induced the unfolding of extracellular proteins, key component of extracellular polymeric substances (EPS), as well as decreased their hydrophobicity and adhesion work by 43.3%, which attenuated the interfacial adhesion of proteins. All the changes caused significant reduction in the colloidal stability and adhesive strength of EPS matrix on the surfaces of films, which eventually disintegrated the biofilms. Consequently, the films achieved over 99.6% inactivation of viable cells and 78.0% reduction of biofilm biomass. This study fabricated a novel antibacterial packaging film that initiates attacks from internal cells to eradicate biofilms.

In neurosurgery, the extent of resection (EOR) is crucial, particularly in the management of malignant gliomas. These tumors are characterized by a diffuse infiltration into the surrounding brain parenchyma. Delineation between tumor and normal brain parenchyma can therefore often be challenging. In recent years, researchers and surgeons have developed and implemented various techniques to improve the visualization of tumors during surgery. This chapter explores current approaches to intraoperative imaging in brain tumor surgery, comprising conventional techniques such as neuronavigation, techniques using fluorescence guided surgery, and further highly precise developments such as targeted-fluorescence, Raman spectroscopy, and hyperspectral imaging. These innovative methods aim to enhance surgeons' ability to identify and remove tumor tissue more effectively, potentially improving patient outcomes.

Optical imaging offers a high potential for noninvasive detection and therapy of cancer in humans. Recent advances in instrumentation for diffuse optical imaging have led to new capabilities for the detection of cancer in highly scattering tissue such as the female breast. In particular, fluorescence imaging is a promising method to image molecular probes in vivo. We review recent developments in the detection of breast cancer and fluorescence-guided surgery of the breast by contrast agents available for application on humans. Detection of cancer has been investigated with the unspecific contrast agents "indocyanine green" and "omocianine" so far. Hereby, indocyanine green was found to offer high potential for the differentiation of malignant and benign lesions by exploiting vessel permeability for macromolecules as a cancer-specific feature. Tumor-specific molecular targeting and activatable probes have been investigated in clinical trials for fluorescence-guided detection of tumor margins. In this application, high spatial resolution can be achieved, since tumor regions are visualized mainly at the tissue surface. As another example of superficial tumor tissue, imaging of lesions in the gastrointestinal tract is discussed. Promising results have been obtained on high-risk patients with Barrett's esophagus and with ulcerative colitis by administering 5-aminolevulinic acid, which induces the accumulation of protoporphyrin IX serving as a tumor-specific fluorescent marker. Time-gated fluorescence imaging and spectroscopy are effective ways to suppress the underlying background from tissue autofluorescence. Furthermore, recently developed tumor-specific molecular probes have been demonstrated to be superior to white-light endoscopy, offering new ways for early detection of malignancies in the gastrointestinal tract.

Dried blood spot (DBS) sampling offers logistical advantages, but it remains unexplored whether analytical results are comparable across DBS sampling methods. This study investigated whether venous or capillary blood spotted on cellulose cards or collected with the Tasso-M20 device provides comparable results for the anti-doping RNA-based biomarkers of erythropoiesis, 5-aminolevulinic acid synthase (ALAS2), and carbonic anhydrase 1 (CA1). In the population of Swiss athletes (n = 12), DBS samples (venous-cellulose and Tasso-M20) were collected in routine anti-doping procedures. In healthy volunteers (n = 12), DBS samples (venous-cellulose, capillary-cellulose and Tasso-M20) were collected at baseline, after 1 week and 10 days post a 3-week recombinant human erythropoietin (rEPO) administration study. ALAS2 and CA1 mRNA were quantified using RT-qPCR, and agreement between matrices was assessed via Passing-Bablok regression. ALAS2 and CA1 expression showed strong linear agreement (r ≥ 0.96) across matrices. Passing-Bablok regression indicated no constant or proportional bias for ALAS2 across all comparisons. For CA1, no bias existed between venous-cellulose and Tasso-M20 in the athlete population, whereas a proportional bias of ~9%-22% was observed when comparing Tasso-M20 DBS with venous- or capillary-cellulose DBS in the rEPO study. No significant differences in relative RNA expression were observed across matrices at any timepoint in the administration study. ALAS2 and CA1 are reliably quantified across venous- and capillary-cellulose-based DBS and Tasso-M20 DBS samples. Strong agreement and minimal bias, with the modest CA1 bias being small relative to expected biological or treatment-induced variability, support their use for longitudinal monitoring, enabling less invasive, flexible, and decentralized sampling in research and anti-doping.

δ-Aminolevulinate dehydratase deficiency porphyria (ADP) is an extremely rare autosomal recessive disorder, with only 12 previously reported cases. It is characterized by the accumulation of delta-aminolevulinic acid (ALA), which is associated with neurovisceral symptoms. We report the first case of ADP in Russia with late-onset symptoms and unique genetic findings. A 47-year-old Russian woman experienced recurrent abdominal pain, tachycardia, hypertension, and neuropathy onset at age 38. Biochemical tests revealed a marked and persistent elevation of urinary ALA, whereas PBG levels showed only moderate and episodic increases. Whole-exome sequencing (WES) revealed two variants in the ALAD gene: a novel heterozygous variant c.299dup, p.(Ala101SerfsTer3) and a mosaic (19% of reads) c.415G>A, p.(Gly139Arg). The c.299dup variant was detected in blood and fibroblasts, while the c.415G>A variant was found only in blood, confirming its mosaic state. The c.415G>A variant was recently reported in a neonatal ADP case as a paternally inherited allele, in contrast with its de novo mosaic origin in our patient. Somatic mosaicism has not been previously described for this disease. Accumulating evidence, including the findings from our case, points to a significant contribution of the erythroid lineage to the disease pathogenesis, suggesting that ADP involves both hepatic and erythroid components, whose relative contributions depend on the tissue-specific genotype and other precipitating factors.

Sustainable biomanufacturing in Escherichia coli is advancing through the integration of carbon dioxide (CO2)-fixing modules from algal systems. However, the assimilation efficiency remains limited by ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) performance and intracellular CO2 availability. To address these constraints, we introduced carbonic anhydrases from Cyanobacteria aponinum (capoCAs) and repressed the native CA (can) gene using Clustered Regularly Interspaced Short Palindromic Repeats interference (CRISPRi) to enhance design specificity. Afterward, capoCAs were overexpressed in E. coli BL21(DE3) and its derivative strains of can deficient (dCA), Calvin-Benson-Bassham (CBB) (RuBisCO-Prk expressing), and dCA+CBB (coupled with dCA and CBB). For functional demonstration, the engineered strains were used to produce a valuable prodrug, 5-aminolevulinic acid (5-ALA). As a result, capoCAs modestly improved 5-ALA titers, with further increments upon CBB coexpression. The optimum dCA+CBB* strain achieved 7.04 g/l (1.46-fold improvement), alongside 9.59% carbon enrichment and a 39.4% reduction in CO2 release. The [13]C-isotope confirmed CO2 incorporation during 5-ALA biosynthesis. This work provides tunable carbon-management modules for developing low-footprint microbial cell factories.

Here, we present a protocol for establishing cutaneous squamous cell carcinoma (cSCC) in SKH-1 hairless mice using a chronic ultraviolet irradiation regimen, followed by treatment with topical 5-aminolevulinic acid photodynamic therapy (ALA-PDT). We describe steps for a 28-week UV induction schedule, standardized preparation and application of ALA cream, and precise parameters for light irradiation. The model recapitulates key stages of human photocarcinogenesis, providing a physiologically relevant system for investigating cSCC pathogenesis and assessing PDT efficacy in vivo. For complete details on the use and execution of this protocol, please refer to Chen et al.[1].

Acute intermittent porphyria (AIP) is a condition characterized by the neurotoxic accumulation of δ-aminolevulinic acid and porphobilinogen. However, chronic kidney disease is an under-recognized complication. A 50-year-old woman experienced recurrent abdominal pain with repeated acute kidney injury and hyponatremia, resulting in a rapid decline in renal function. Marked elevations of both metabolites, as well as her genetic mutation, confirmed AIP. The administration of hemin markedly decreased the frequency of porphyria attacks and significantly attenuated the annual estimated glomerular filtration rate decline. Although the patient eventually developed end-stage kidney disease, hemin treatment slowed the progression of renal failure.

This study compared apoptosis induction in squamous cell carcinoma keratinocytes following exposure to 5-aminolevulinic acid combined with either blue light (417 ± 5 nm) or red light (635 ± 5 nm) photodynamic therapy, with apoptotic activity quantified via levels of Annexin V expression. Our results demonstrated that blue light induces superior levels of apoptosis in squamous cell carcinoma keratinocytes compared to red light at the parameters tested. Both wavelengths demonstrate additional dose- and temperature-dependent effects across two squamous cell carcinoma cell lines. These conclusions had significant implications for the clinical selection of a light source in combination with 5-aminolevulinic acid photodynamic therapy in the treatment of actinic keratosis and early squamous cell carcinoma lesions.

In our previous study, 5-aminolevulinic acid-mediated photodynamic therapy (ALA-PDT) promoted chronic diabetic wound healing by reducing the microbial burden. However, the anti-bacterial efficacy of ALA-PDT is often compromised by limited light penetration into tissues. To increase the efficacy of ALA-PDT, this study assessed its combined use with the antibiotic ceftazidime in treating chronic diabetic wounds and examined the potential mechanism of action. The results indicated that the combination of ALA-PDT and ceftazidime synergistically inhibited the growth of multidrug-resistant Pseudomonas aeruginosa. The treatment also prevented the formation of bacterial biofilm, delayed the development of antibiotic resistance, inactivated pyocyanin, and exerted a prolonged post-antibiotic effect. The mechanism supporting the synergistic antibacterial effect might involve ALA-PDT-mediated reactive oxygen species generation, leading to bacterial cell structure destruction, increased membrane permeability, and cellular ceftazidime uptake. In a full-thickness skin defect infection model in rats with diabetes, the bactericidal and wound closure rates were highest in the combination therapy group. Transcriptomics analysis of wound tissues revealed that the differentially expressed genes between the combination therapy and model groups were mainly enriched in the IL-17, AKT-STAT, and VEGF signaling pathways. Molecular biology experiments illustrated that the combination therapy might regulate the expression of IL-17, activate the JAK-STAT axis, increase p-JAK2 and p-STAT3 expression, and decrease SOCS3 expression, ultimately leading to Ki-67, CD31, and VEGF upregulation and exerting beneficial effects on angiogenesis, collagen deposition, and tissue regeneration. Overall, PDT and antibiotic combination therapy offers an efficient approach to treat infections in diabetic wounds.

Basal cell carcinoma (BCC) is a common skin malignancy, often occurring in cosmetically sensitive facial areas. We report a case of a 46-year-old woman with BCC of the right nasal ala. The lesion had persisted for nine years, gradually enlarging, with recent ulceration and crusting. Skin biopsy confirmed BCC. The patient underwent Mohs micrographic surgery (MMS), with the wound left to heal by secondary intention for 48 h, followed by sequential 5-aminolevulinic acid photodynamic therapy (ALA-PDT) once weekly for five consecutive weeks. A freshly prepared 20 % 5-aminolevulinic acid (ALA) solution (118 mg, Shanghai Fudan-Zhangjiang Bio-Pharmaceutical Co., Ltd) was topically applied to the entire open defect of the right nasal ala and its 0.5 cm peripheral marginal area, occluded with aluminum foil to block light, and incubated for 3 h. After removal of the covering, the area was irradiated with 635-nm red light at 80 mW/cm[2]and 120 J/cm[2] for 20 min (Wuhan Yage Optic and Electronic Technique Co., Ltd). Approximately 5 months and 12 days after the final PDT session, repeat histopathology showed no residual tumor, and hypertrophic scar formation was observed. No recurrence or further BCC was noted during the 5-year follow-up. This case demonstrates that MMS combined with ALA-PDT can achieve effective local tumor control while preserving function and optimizing cosmetic outcome for BCCs larger than 1 cm in aesthetically sensitive facial areas.