Expression of human Ras-related protein Rab39B variant T168K in Caenorhabditis elegans leads to motor dysfunction and dopaminergic neuron degeneration.

Human RAB39B gene is related to familial early-onset Parkinson disease. In early adulthood, men with the RAB39B c.503C > A (Thr168Lys, p. T168K) mutation develop typical tremor, bradykinesia, and alpha-synuclein accumulation. We investigated the pathological mechanism of RAB39B T168K in a Caenorhabditis elegans model. In early adult C. elegans, RAB39B T168K led to dopaminergic neuron degeneration that presented as disrupted dendrites and blunt neuronal cells. Abnormal dopamine secretion was inferred from a decline in motor function and a positive basal slowing phenotype. Dopamine-associated tests confirmed that synthesis and recycling of dopamine were normal. The RAB39B T168K mutation might impair dopamine vesicular transmission from the presynaptic membrane to the synaptic gap in dopaminergic neurons. The release-dependent feedback mechanism in neurotransmitters regulates the balance of receptor activities. Protein-protein interactions network analysis revealed that RAB39B may also function in lysosomal degradation and autophagy. Impaired disposal of misfolded α-synuclein eventually leads to protein aggregation. Thus, like other members of the Rab family, RAB39B may be involved in vesicular transport associated with dopamine secretion and α-synuclein clearance.

Cortical microinfarcts potentiate recurrent ischemic injury through NLRP3-dependent trained immunity.

Microinfarcts are common among the elderly and patients with microinfarcts are more vulnerable to another stroke. However, the impact of microinfarcts on recurrent stroke has yet to be fully understood. The purpose of this study was to explore the negative effects of microinfarcts on recurrent stroke. To achieve this, two-photon laser was used to induce microinfarcts, while photothrombotic stroke was induced on the opposite side. The results showed that microinfarcts led to trained immunity in microglia, which worsened the pro-inflammatory response and ischemic injury in the secondary photothrombotic stroke. Additionally, the study clarified the role of NLRP3 in microglial nuclei, indicating that it interacts with the MLL1 complex through NACHT domain and increases H3K4 methylation, which suggests that NLRP3 is critical in the formation of innate immune memory caused by microinfarcts. Furthermore, the knockout of NLRP3 in microglia alleviated the trained immunity and reduced the harmful effects of microinfarcts on recurrent stroke. This study emphasizes the detrimental effect of trained immunity on recurrent stroke and highlights the critical role of NLRP3 in mediating the formation of this memory, which may offer a potential therapeutic target for mitigating recurrent strokes.

Phosphorylation of AQP4 by LRRK2 R1441G impairs glymphatic clearance of IFNγ and aggravates dopaminergic neurodegeneration.

Aquaporin-4 (AQP4) is essential for normal functioning of the brain's glymphatic system. Impaired glymphatic function is associated with neuroinflammation. Recent clinical evidence suggests the involvement of glymphatic dysfunction in LRRK2-associated Parkinson's disease (PD); however, the precise mechanism remains unclear. The pro-inflammatory cytokine interferon (IFN) γ interacts with LRRK2 to induce neuroinflammation. Therefore, we examined the AQP4-dependent glymphatic system's role in IFNγ-mediated neuroinflammation in LRRK2-associated PD. We found that LRRK2 interacts with and phosphorylates AQP4 in vitro and in vivo. AQP4 phosphorylation by LRRK2 R1441G induced AQP4 depolarization and disrupted glymphatic IFNγ clearance. Exogeneous IFNγ significantly increased astrocyte expression of IFNγ receptor, amplified AQP4 depolarization, and exacerbated neuroinflammation in R1441G transgenic mice. Conversely, inhibiting LRRK2 restored AQP4 polarity, improved glymphatic function, and reduced IFNγ-mediated neuroinflammation and dopaminergic neurodegeneration. Our findings establish a link between LRRK2-mediated AQP4 phosphorylation and IFNγ-mediated neuroinflammation in LRRK2-associated PD, guiding the development of LRRK2 targeting therapy.

Real-world experience with Deutetrabenazine management in patients with Huntington's disease using video-based telemedicine.

BACKGROUND: Huntington's disease (HD) is a rare progressive neurological disorder, and telemedicine has the potential to improve the quality of care for patients with HD. Deutetrabenazine (DTBZ) can reduce chorea symptoms in HD; however, there is limited experience with this medication in Asian countries. METHODS: Retrospective and prospective studies were employed to explore the feasibility and reliability of a video-based telemedicine system for HD patient care. Reliability was demonstrated through consistency between selected-item scores (SIS) and total motor scores (TMS) and the agreement of scores obtained from hospital and home videos. Finally, a single-centre real-world DTBZ management study was conducted based on the telemedicine system to explore the efficacy of DTBZ in patients with HD. RESULTS: There were 77 patients included in the retrospective study, and a strong correlation was found between SIS and TMS (r = 0.911, P < 0.0001), indicating good representativeness. There were 32 patients enrolled in the prospective study. The reliability was further confirmed, indicated by correlations between SIS and TMS (r = 0.964, P < 0.0001) and consistency of SIS derived from the in-person and virtual visits (r = 0.969, P < 0.0001). There were 17 patients included in the DTBZ study with a mean 1.41 (95% confidence interval, 0.37-2.46) improvement in chorea score and reported treatment success. CONCLUSIONS: A video-based telemedicine system is a feasible and reliable option for HD patient care. It may also be used for drug management as a supplementary tool for clinical visits.

Divergent Synthesis of Chromenoindoles and Spiroindolines via Domino Reaction of Indolyl-Substituted Isocyanides with Quinone Esters.

A dearomative spirocyclization of tryptamine-derived isocyanides with quinone esters is developed for the divergent synthesis of structurally complex chromeno[2,3-b]indole and polycyclic spiroindoline scaffolds. This domino reaction features the formation and conversion of the six-membered dihydropyran ring with an amendable N,O-aminal moiety.

Evaluation of Blood Glial Fibrillary Acidic Protein as a Potential Marker in Huntington's Disease.

Objective: Huntington's disease (HD) is an autosomal dominant neurodegenerative disorder. Neurofilament light protein (NfL) is correlated with clinical severity of HD but relative data are the lack in the Chinese population. Reactive astrocytes are related to HD pathology, which predicts their potential to be a biomarker in HD progression. Our aim was to discuss the role of blood glial fibrillary acidic protein (GFAP) to evaluate clinical severity in patients with HD. Methods: Fifty-seven HD mutation carriers (15 premanifest HD, preHD, and 42 manifest HD) and 26 healthy controls were recruited. Demographic data and clinical severity assessed with the internationally Unified Huntington's Disease Rating Scale (UHDRS) were retrospectively analyzed. Plasma NfL and GFAP were quantified with an ultra-sensitive single-molecule (Simoa, Norcross, GA, USA) technology. We explored their consistency and their correlation with clinical severity. Results: Compared with healthy controls, plasma NfL (p < 0.0001) and GFAP (p < 0.001) were increased in Chinese HD mutation carriers, and they were linearly correlated with each other (r = 0.612, p < 0.001). They were also significantly correlated with disease burden, Total Motor Score (TMS) and Total Functional Capacity (TFC). The scores of Stroop word reading, symbol digit modalities tests, and short version of the Problem Behaviors Assessments (PBAs) for HD were correlated with plasma NfL but not GFAP. Compared with healthy controls, plasma NfL has been increased since stage 1 but plasma GFAP began to increase statistically in stage 2. Conclusions: Plasma GFAP was correlated with plasma NfL, disease burden, TMS, and TFC in HD mutation carriers. Plasma GFAP may have potential to be a sensitive biomarker for evaluating HD progression.

Therapeutic reversal of Huntington's disease by in vivo self-assembled siRNAs.

Huntington's disease is an autosomal-dominant neurodegenerative disease caused by CAG expansion in exon 1 of the huntingtin (HTT) gene. Since mutant huntingtin (mHTT) protein is the root cause of Huntington's disease, oligonucleotide-based therapeutic approaches using small interfering RNAs (siRNAs) and antisense oligonucleotides designed to specifically silence mHTT may be novel therapeutic strategies for Huntington's disease. Unfortunately, the lack of an effective in vivo delivery system remains a major obstacle to realizing the full potential of oligonucleotide therapeutics, especially regarding the delivery of oligonucleotides to the cortex and striatum, the most severely affected brain regions in Huntington's disease. In this study, we present a synthetic biology strategy that integrates the naturally existing exosome-circulating system with artificial genetic circuits for self-assembly and delivery of mHTT-silencing siRNA to the cortex and striatum. We designed a cytomegalovirus promoter-directed genetic circuit encoding both a neuron-targeting rabies virus glycoprotein tag and an mHTT siRNA. After being taken up by mouse livers after intravenous injection, this circuit was able to reprogramme hepatocytes to transcribe and self-assemble mHTT siRNA into rabies virus glycoprotein-tagged exosomes. The mHTT siRNA was further delivered through the exosome-circulating system and guided by a rabies virus glycoprotein tag to the cortex and striatum. Consequently, in three mouse models of Huntington's disease treated with this circuit, the levels of mHTT protein and toxic aggregates were successfully reduced in the cortex and striatum, therefore ameliorating behavioural deficits and striatal and cortical neuropathologies. Overall, our findings establish a convenient, effective and safe strategy for self-assembly of siRNAs in vivo that may provide a significant therapeutic benefit for Huntington's disease.

Transmembrane Protein 230 Mediates a Poly(ADP-ribose) Polymerase-1-Linked Apoptosis.

Mutations in transmembrane protein 230 (TMEM230) gene are suggested to be associated with the autosomal dominant Parkinson's disease (PD) with typical movement disorders and Lewy body pathology. However, the normal functions and the pathological roles of TMEM230 are not clear. In this study, we used TMEM230 isoform II constructs including wild-type (WT) and four reported PD-linked mutation constructs (Y92C, R141L, 184Wext*5, and 184PGext*5). Ectopic expression of WT and PD-linked mutant TMEM230 variants in cultured cells dramatically induced apoptotic cell death compared with that of vector control cells. Mutant TMEM230 caused cell toxicity at an increased severity than WT TMEM230. Moreover, expression of TMEM230 increased mitochondrial reactive oxygen species (ROS) levels, decreased cellular ATP, activated caspase 3/7, and increased poly(ADP-ribose) polymerase-1 (PARP1) cleavage. Treatment with N-acetylcysteine (NAC; an ROS scavenger) or Z-VAD-FMK (a caspase inhibitor) significantly attenuated TMEM230-induced apoptosis in both cultured cells and primary neurons. Our results indicated that TMEM230 mediated a PARP1-linked apoptotic cell death pathway. These findings not only provide the novel insight into the biological roles of TMEM230 in the PARP1-linked pathway but also provide a TMEM230-induced cell death mechanism underlying PD pathogenesis.

Mesenchymal stem cell-derived exosomes improve motor function and attenuate neuropathology in a mouse model of Machado-Joseph disease.

BACKGROUND: Machado-Joseph disease is the most common autosomal dominant hereditary ataxia worldwide without effective treatment. Mesenchymal stem cells (MSCs) could slow the disease progression, but side effects limited their clinical application. Besides, MSC-derived exosomes exerted similar efficacy and have many advantages over MSCs. The aim of this study was to examine the efficacy of MSC-derived exosomes in YACMJD84.2 mice. METHODS: Rotarod performance was evaluated every 2 weeks after a presymptomatic administration of intravenous MSC-derived exosomes twice in YACMJD84.2 mice. Loss of Purkinje cells, relative expression level of Bcl-2/Bax, cerebellar myelin loss, and neuroinflammation were assessed 8 weeks following treatment. RESULTS: MSC-derived exosomes were isolated and purified through anion exchange chromatography. Better coordination in rotarod performance was maintained for 6 weeks in YACMJD84.2 mice with exosomal treatment, compared with those without exosomal treatment. Neuropathological changes including loss of Purkinje cells, cerebellar myelin loss, and neuroinflammation were also attenuated 8 weeks after exosomal treatment. The higher relative ratio of Bcl-2/Bax was consistent with the attenuation of loss of Purkinje cells. CONCLUSIONS: MSC-derived exosomes could promote rotarod performance and attenuate neuropathology, including loss of Purkinje cells, cerebellar myelin loss, and neuroinflammation. Therefore, MSC-derived exosomes have a great potential in the treatment of Machado-Joseph disease.

Electrocatalysis of N2 to NH3 by HKUST-1 with High NH3 Yield.

Electrolytic ammonia synthesis from nitrogen at ambient conditions is appearing as a promising alternative to the Haber-Bosch process which is consuming high energy and emitting CO2 . Here, a typical MOF material, HKUST-1 (Cu-BTC, BTC=benzene-1,3,5-tricarboxylate), was selected as an electrocatalyst for the reaction of converting N2 to NH3 under ambient conditions. At -0.75 V vs. reversible hydrogen electrode, it achieves excellent catalytic performance in the electrochemical synthesis of ammonia with high NH3 yield (46.63 µg h-1 mg-1 cat. or 4.66 µg h-1 cm-2 ) and good Faraday efficiency (2.45%). It is indicated that the good performance of the HKUST-1 catalyst may originate from the formation of Cu(I). In addition, the catalyst also has good selectivity for N2 to NH3 .

Mesenchymal stem cells alleviate AQP-4-dependent glymphatic dysfunction and improve brain distribution of antisense oligonucleotides in BACHD mice.

Huntington's disease (HD) is a neurodegenerative disorder caused by a mutation in the huntingtin (HTT) gene that results in the production of neurotoxic mutant HTT (mHTT) protein. Suppressing HTT production with antisense oligonucleotides (ASOs) is a promising treatment strategy for HD; however, the difficulty of delivering ASOs to deep brain structures is a major barrier for its clinical application. The glymphatic system of astrocytes involving aquaporin 4 (AQP-4) controls the entry of macromolecules from the cerebrospinal fluid into the brain. Mesenchymal stem cells (MSCs) target astrocytes to inhibit neuroinflammation. Here we examined the glymphatic distribution of ASO in the brain and the therapeutic potential of combining intravenously injection of mesenchymal stem cells (IV-MSC) and ASOs for the treatment of HD. Our results show that Cy3-labeled ASOs entered the brain parenchyma via the perivascular space following cisternal injection, but the brain distribution was significantly lower in AQP-4-/- as compared with wild-type mice. Downregulation of the AQP-4 M23 isoform was accompanied by decreased brain levels of ASOs in BACHD mice as well as an increase in astrogliosis and phosphorylation of nuclear factor κB (NF-κB) p65. IV-MSC treatment restored AQP-4 M23 expression, attenuated astrogliosis, and decreased NF-κB p65 phosphorylation; it also increased the brain distribution of ASOs and enhanced the suppression of mHTT in BACHD mice. These results suggest that modulating glymphatic activity using IV-MSC is a novel strategy for improving the potency of ASO in the treatment of HD.

Liangyi Gao extends lifespan and exerts an antiaging effect in Caenorhabditis elegans by modulating DAF-16/FOXO.

Liangyi Gao (LYG), a traditional Chinese medicine, is composed of Ginseng and Radix Rehmanniae Preparata, both of which have been shown to have antiaging properties. In Eastern countries, LYG is used to delay functional declines related to aging and has an obvious antiaging effect in clinical practice. However, little data from evidence-based medicine is available regarding whether LYG is beneficial overall, particularly with respect to lifespan, and how LYG functions. To address these issues, Caenorhabditis elegans, a useful organism for such studies, was employed to explore the antiaging effect and mechanism of LYG in this study. The results showed that LYG could obviously extend lifespan and slow aging-related declines in N2 wild-type C. elegans. To further characterize these antiaging effects and stress resistance, reproductive tests and other aging-related tests were performed. We found that LYG enhanced resistance against oxidative and thermal stress, reproduction, pharynx pumping, motility and growth in N2 wild-type C. elegans. In addition, we analyzed the mechanism for these effects by measuring the activity of superoxide dismutase (SOD) and the expression levels of aging-related genes. We found that LYG enhanced the activities of antioxidant enzymes and upregulated the genes daf-16, sod-3 and sir-2.1, which mediated stress resistance and longevity. In conclusion, LYG had robust and reproducible life-prolonging and antiaging benefits in C. elegans via DAF-16/FOXO regulation.

A dual (colorimetric and fluorometric) detection scheme for glutathione and silver (I) based on the oxidase mimicking activity of MnO2 nanosheets.

A fluorimetric and colorimetric method is described for the determination of glutathione (GSH) and silver (I). It is based on the use of MnO2 nanosheets that were prepared by solution mixing and exfoliation. They display oxidase-mimicking activity and can catalyze the oxidation of o-phenylenediamine (OPD) to form yellow 2,3-diaminophenazine (DAP) with an absorption maximum at 410 nm. DAP also has a yellow fluorescence (with a peak at 560 nm). The MnO2 nanosheets can be rapidly reduced to Mn2+ by GSH. This reduces the efficiency of the oxidase mimic MnO2 and causes a decrease in fluorescence and absorbance intensity. However, on addition of Ag+, a complex is formed with GSH. It prevents the destruction of MnO2 nanosheets so that the enzyme mimicking activity is retained. A dual-method for the determination of GSH and Ag(I) was developed. It has excellent sensitivity for GSH with lower detection limits of 62 nM (fluorimetric) and 0.94 µM (colorimetric). The respective data for Ag(I) are 70 nM and 1.15 µM. The assay was successfully applied to the determination of GSH and Ag(I) in spiked serum samples. Graphical abstract Schematic presentation of a method for colorimetric and fluorometric determination of glutathione (GSH) and silver(I). MnO2 nanosheets are reduced to Mn(II) by GSH. This reduces the enzyme-mimicking activity of MnO2 nanosheets and causes a decrease in fluorescence and absorbance. On addition of Ag(I), the enzyme-like activity is increasingly retained. A decrease in fluorescence and absorbance is not observed any longer.

Colorimetric detection of ascorbic acid and alkaline phosphatase activity based on the novel oxidase mimetic of Fe-Co bimetallic alloy encapsulated porous carbon nanocages.

A novel catalyst of FeCo nanoparticles (FeCo NPs) incorporated porous nanocages (FeCo NPs@PNC) was first synthesized by encapsulating of FeCo alloy into ZIF-8 and further carbonation of the composite. The FeCo NPs@PNC displays enhanced intrinsic oxidase-like activity compared to the individual FeCo NPs and porous nanocages (PNC). The FeCo NPs@PNC can catalyze the oxidation of 3,3',5,5'-tetramethylbenzidine (TMB) to oxidized TMB (oxTMB) without H2O2, producing a blue color with a maximum absorption peak at 652 nm. The catalytic mechanism was investigated and it found that the intermediate (O2·-) produced from the catalytic process in the system of TMB-O2-FeCo NPs@PNC can accelerate the oxidation of TMB to oxTMB. However, ascorbic acid (AA) can reduce the oxTMB and result in a conspicuous blue color fading. Therefore, a novel colorimetric platform was constructed to quantify AA with the linear range of 0.5-28 µM and detection limit of 0.38 µM (at 3σ/m). Owing to the alkaline phosphatase (ALP) can catalyze the hydrolysis of AA 2-phosphate (AAP) into AA, ALP can also be quantified by the above method. And the linear range for ALP is 0.6-10 U L-1 and the limit of detection is 0.49 U L-1. The FeCo NPs@PNC also shows excellent stability and reproducibility. This study provides a new alternative oxidase mimetic on the basis of easily obtained metal-organic frameworks derivatives to replace the expensive natural enzymes and noble metal based nanoenzymes, which will show great potential in biological assays.

Dual II Heterojunctions Metallic Phase MoS2 /ZnS/ZnO Ternary Composite with Superior Photocatalytic Performance for Removing Contaminants.

Fabricating a high-performance photocatalyst to efficiently solve serious environmental problems is an urgent affair. Herein, a series of MoS2 /ZnO composites were successfully fabricated through a facile hydrothermal route using Na2 MoO4 , (NH2 )2 CS and urchin-like ZnO as precursors. According to the results of XRD and XPS, it was found that ZnS appeared in MoS2 /ZnO composite; meanwhile, the content was positively correlated with the weight of the precursor (NH2 )2 CS. It should be noted that the morphology and the metallic phase content of MoS2 grown in situ on the surface of ZnO were affected by the molar ratio of Na2 MoO4 and ZnO. Benefiting from the special dual II heterojunctions of MoS2 /ZnS/ZnO ternary composite, the material exhibited excellent charge separation and transfer performances. In the photocatalytic measurements, the MoS2 /ZnS/ZnO (Na2 MoO4 :ZnO 1:2 MZ2) composite not only exhibits excellent photocatalytic CrVI reduction activity of 42.3×10-3  min-1 , but also displays remarkable adsorption performance (nearly 32.1 %) for Cr2 O 2 - 7 . In addition, the ternary composite shows dominant photocatalytic CrVI reduction activities compared to other photocatalysts. This work provides a high-efficient MoS2 /ZnS/ZnO ternary photocatalyst for environmental treatment.

A sensitive electrochemical sensor for bisphenol A on the basis of the AuPd incorporated carboxylic multi-walled carbon nanotubes.

A sensitive electrochemical sensor for BPA based on the AuPd incorporated carboxylic multi-walled carbon nanotubes (MWCNT) with synergetic amplified current signal was developed, where MWCNT was used as supporter to improve electron transport and poly (diallyldimethylammonium chloride) (PDDA) was used as dispersant for MWCNT and overcome the intrinsic van der Waals interactions between MWCNT and further increase metal NPs loading. The prepared MWCNT-PDDA-AuPd showed enhanced electrocatalytic performance toward BPA, which is better than those of homologous monometallic counterparts and MWCNT-PDDA though the content of AuPd is really low. The peak currents of BPA increased with BPA concentration in linear range of 0.18-18 µM and the detection limit of 60 nM. The sensor showed high sensitivity, good stability, repeatability and can be used to detect BPA in milk and water samples with good performance, which demonstrate that MWCNTs-PDDA-AuPd nanocomposite may be an attractive material in applications of environmental and food analysis.

Biomass-derived oxygen-doped hollow carbon microtubes for electrocatalytic N2-to-NH3 fixation under ambient conditions.

Electrocatalytic N2 reduction as an alternative approach to the energy-intensive and large CO2-producing Haber-Bosch process for NH3 synthesis under mild conditions has attracted extensive attention. Current research efforts on N2 reduction have mainly focused on metal-based catalysts, but metal-free alternatives can avoid the issue of metal ion release. In this work, oxygen-doped hollow carbon microtubes (O-KFCNTs) derived from natural kapok fibers are reported as a metal-free NRR electrocatalyst for N2-to-NH3 conversion with excellent selectivity. In 0.1 M HCl, the O-KFCNTs achieve a high faradaic efficiency of 9.1% at -0.80 V vs. a reversible hydrogen electrode (RHE) and a NH3 yield rate of 25.12 µg h-1 mgcat.-1 at -0.85 V vs. RHE under ambient conditions. Notably, this catalyst also demonstrates high stability.

A novel biomimetic nanoenzyme based on ferrocene derivative polymer NPs coated with polydopamine.

In this paper, ferrocene derivative polymer nanoparticles (FcP NPs) with uniform size and good photostability was synthesized using 1,1'-ferrocene dicarboxylic acid as precursor and methanol as solvent. FcP NPs-PDA was further obtained by coating of polydopamine (PDA) on FcP NPs in tris-HCl (pH=8.5) buffer solution at room temperature in the presence of dopamine (DA). The structure and morphology of FcP NPs and FcP NPs-PDA were characterized by transmission electron microscope (TEM), ultraviolet-visible spectroscopy (UV-Vis), and infrared radiation (FT-IR). The as-prepared FcP NPs-PDA showed better peroxidase-like activity than FcP NPs, which could catalyze the chromogenic reaction of peroxidase substrate TMB, OPD and ABTS. Based on the high peroxidase-like property of FcP NPs-PDA, a sensitive and convenient means to detect H2O2 has been proposed with TMB as the substrate, which displays wide linear range of 10-600 µM and 600 µM-4 mM with low detection limit of 5 µΜ. Compared with other Fe-containing NPs, such as magnetic materials and noble metal@Fe bimetallic NPs, the preparation approach for FcP NPs-PDA is simple, time and energy saving and environment friendly. This mild and simple synthesis route of FcP NPs-PDA will provide new ideas for the preparation of non-noble metal-based peroxidase-like nanomaterials and widen the applications in the fields of catalytic and analytical chemistry.

Colorimetric determination of ascorbic acid and the activity of alkaline phosphatase based on the inhibition of the peroxidase-like activity of citric acid-capped Prussian Blue nanocubes.

Colorimetric methods are described for the determination of ascorbic acid (AA) and alkaline phosphatase (ALP). Both assays are based on the inhibition of the peroxidase (POx)-like activity of Prussian Blue nanocubes (PB NCs) capped with citric acid. They catalyze the oxidation of 3,3,5,5-tetramethylbenzidine (TMB) by H2O2 to produce a blue color with an absorption maximum at 652 nm. On addition of AA, the PB NCs are reduced to Prussian White (PW) which does not act as a POx mimic. This results in a decreased rate of the formation of the blue coloration whose intensity decreases with increasing concentration of AA. The assay allows AA to be quantified with a 35 nM detection limit (at 3σ/m). The hydrolysis of AA phosphate by ALP leads to the formation of AA which can be quantified by the above method. Based on this, the activity of ALP can be determined by measurement of the intensity of the blue coloration thus formed. The method can be used to determine the activity of ALP with a detection limit as low as 0.23 U·L-1. Graphical abstract Schematic presentation of a method for colorimetric determination of ALP activity. AA obtained by ALP-catalyzed hydrolysis of ascorbic acid phosphate (AAP) inhibits the intrinsic peroxidase-like activity of PB NCs by reducing Prussian Blue nanocrystals (PB NCs) to form inactive Prussian White (PW).