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Lab on a Chip Jan 2024Multiple protocols have been reported to fabricate paper-based analytical devices (PADs). However, some of these techniques must be revised because of the...
Multiple protocols have been reported to fabricate paper-based analytical devices (PADs). However, some of these techniques must be revised because of the instrumentation required. This paper describes a versatile and globally affordable method to fabricate PADs using office paper as a substrate and a laser printing technique to define hydrophobic barriers on paper surfaces. To demonstrate the feasibility of the alternatives proposed in this study, the fabrication of devices for three types of detection commonly associated with using PADs was demonstrated: colorimetric detection, electrochemical detection, and mass spectrometry associated with a paper-spray ionization (PSI-MS) technique. Besides that, an evaluation of the type of paper used and chemical modifications required on the substrate surface are also presented in this report. Overall, the developed protocol was suitable for using office paper as a substrate, and the laser printing technique as an efficient fabrication method when using this substrate is accessible at a resource-limited point-of-need. Target analytes were used as a proof of concept for these detection techniques. Colorimetric detection was carried out for acetaminophen, iron, nitrate, and nitrite with limits of detection of 0.04 μg, 4.5 mg mL, 2.7 μmol L, and 6.8 μmol L, respectively. A limit of detection of 0.048 fg mL was obtained for the electrochemical analysis of prostate-specific antigen. Colorimetric and electrochemical devices revealed satisfactory performance when office paper with a grammage of 90 g m was employed. Methyldopa analysis was also carried out using PSI-MS, which showed a good response in the same paper weight and behavior compared to chromatographic paper.
PubMed: 38126917
DOI: 10.1039/d3lc00840a -
The Ocular Surface Jan 2024Herpes simplex keratitis (HSK), caused by type 1 herpes simplex virus (HSV) reactivation, is a severe infectious disease that leads to vision loss. HSV can trigger...
PURPOSE
Herpes simplex keratitis (HSK), caused by type 1 herpes simplex virus (HSV) reactivation, is a severe infectious disease that leads to vision loss. HSV can trigger metabolic reprogramming in the host cell and change the extracellular vesicles (EV) cargos; however, little is known about the EV metabolic signatures during ocular HSV infection. Here, we aimed to depict the EV-associated metabolic landscape in HSK patients' tears.
METHODS
We collected 82 samples from 41 participants with unilateral HSK (contralateral unaffected tears were set as negative control), including subtype cohorts of 13 epithelial, 20 stromal, and 8 endothelial HSK. We isolated tear EVs via our previously established platform and conducted metabolic analysis using LC-MS/MS. The metabolic signatures for recognizing HSK and subtypes were assessed through differential analysis and machine learning algorithms.
RESULTS
Hypopsia and increased extracellular CD63 levels were observed in affected eyes. We identified 339 metabolites based on sEVs isolated from tears. Differential analysis revealed alterations in energy and amino acid metabolism, as well as the infectious microenvironment. Furthermore, we observed dysregulated metabolite such as methyldopa, which is associated with inappropriate neovascularization and corneal sensation loss, contributing to the HSK severity particularly in the stromal subtype. Moreover, machine learning classification also suggested a set of EV metabolic signatures that have potential for pan-keratitis detection.
CONCLUSIONS
Our findings demonstrate that tear EV metabolites can serve as valuable indicators for comprehending the underlying pathological mechanisms. This knowledge is expected to facilitate the development of liquid biopsy means and therapeutic target discovery.
Topics: Humans; Chromatography, Liquid; Tandem Mass Spectrometry; Keratitis, Herpetic; Cornea; Simplexvirus
PubMed: 38122863
DOI: 10.1016/j.jtos.2023.12.005 -
Frontiers in Neurology 2023Aromatic L-amino acid decarboxylase (AADC) deficiency is a rare autosomal recessive neurometabolic disorder leading to severe combined serotonin, dopamine,...
Case report: Childhood epilepsy and borderline intellectual functioning hiding an AADC deficiency disorder associated with compound heterozygous gene pathogenic variants.
Aromatic L-amino acid decarboxylase (AADC) deficiency is a rare autosomal recessive neurometabolic disorder leading to severe combined serotonin, dopamine, norepinephrine, and epinephrine deficiency. We report on a female patient with borderline functioning and sporadic clear-cut focal to bilateral seizures from age 10 years. A neuropsychological assessment highlighted a mild impairment in executive functions, affecting attention span and visual-spatial abilities. Following the diagnosis of epilepsy with a presumed genetic etiology, we applied a diagnostic approach inclusive of a next-generation sequencing (NGS) gene panel, which uncovered two variants in the DOPA decarboxylase () gene underlying an AADC deficiency. This compound heterozygous genotype was associated with a mild reduction of homovanillic acid, a low level of the norepinephrine catabolite, and a significant reduction of 5-hydroxyindoleacetic acid in cerebrospinal fluid. Remarkably, 3-O-methyldopa (3-OMD) and 5-hydroxytryptophan were instead increased. During the genetically guided re-evaluation process, some mild signs of dysautonomic dysfunction (nasal congestion, abnormal sweating, hypotension and fainting, excessive sleepiness, small hands and feet, and increased levels of prolactin, tiredness, and fatigue), more typical of AADC deficiency, were evaluated with new insight. Of the two AADC variants, the R347Q has already been characterized as a loss-of-function with severe catalytic impairments, while the novel L391P variant has been predicted to have a less severe impact. Bioinformatic analyses suggest that the amino acid substitution may affect affinity for the PLP coenzyme. Thus, the genotype corresponds to a phenotype with mild and late-onset symptoms, of which seizures were the clinical sign, leading to medical attention. This case report expands the spectrum of AADC deficiency phenotypes to encompass a less-disabling clinical condition including borderline cognitive functioning, drug-responsive epilepsy, and mild autonomic dysfunction.
PubMed: 38116105
DOI: 10.3389/fneur.2023.1284339 -
Movement Disorders : Official Journal... Feb 2024IPX203 is a novel oral extended-release formulation of carbidopa/levodopa (CD/LD) developed to address the short half-life of immediate-release CD/LD. In the phase 3... (Randomized Controlled Trial)
Randomized Controlled Trial
BACKGROUND
IPX203 is a novel oral extended-release formulation of carbidopa/levodopa (CD/LD) developed to address the short half-life of immediate-release CD/LD. In the phase 3 RISE-PD trial, IPX203 significantly improved "Good On" time in patients with Parkinson's disease compared with immediate-release CD/LD.
OBJECTIVES
To evaluate the safety and efficacy of IPX203 in an open-label extension of the pivotal phase 3 study.
METHODS
This 9-month extension enrolled patients who completed the randomized, double-blind trial. Key efficacy endpoints included Movement Disorder Society-Unified Parkinson's Disease Rating Scale and Patient and Clinical Global Impression scores. Adverse events (AEs) were recorded.
RESULTS
Improvements in efficacy were maintained and dosing frequency and total daily dose remained stable through the trial. A total of 52.7% of patients experienced ≥1 treatment-emergent AE, mostly mild or moderate and occurred within the first 90 days of treatment.
CONCLUSIONS
In this phase 3 open-label extension, IPX203 exhibited a favorable safety and tolerability profile and sustained efficacy of comparable magnitude to the end of the double-blind study. © 2023 The Authors. Movement Disorders published by Wiley Periodicals LLC on behalf of International Parkinson and Movement Disorder Society.
Topics: Humans; Parkinson Disease; Antiparkinson Agents; Levodopa; Carbidopa; Delayed-Action Preparations; Research; Drug Combinations; Double-Blind Method
PubMed: 38111267
DOI: 10.1002/mds.29685 -
Rinsho Shinkeigaku = Clinical Neurology Jan 2024A 70-year-old male who has medical history of Parkinson's disease for 26 years admitted to our hospital for trial of levodopa carbidopa intestinal gel (LCIG) therapy...
[A 70-year-old male exhibiting parkinsonism-hyperpyrexia syndrome during levodopa carbidopa intestinal gel (LCIG) treatment and suspected carbidopa allergy due to positive drug-induced lymphocyte stimulation test (DLST)].
A 70-year-old male who has medical history of Parkinson's disease for 26 years admitted to our hospital for trial of levodopa carbidopa intestinal gel (LCIG) therapy because of severe dyskinesia and frequent wearing-off. He developed deterioration when he was treated with one of the levodopa (LD) decacrboxylase inhibitor compounds in the past. Five days after LD had changed into equivalent dose of LD/carbidopa (CD), high fever with hyperCKemia appeared. He was diagnosed as having Parkinsonism-hyperpyrexia syndrome (PHS). Exchange of LD/CD to LD drugs improved the symptoms quickly. Four days after LCIG administration, PHS reappeared. Simultaneously, the patient developed sepsis and disseminated intravascular coagulation (DIC). Thrombocytopenia did not improve after recovery from infection and DIC. Anti-PA IgG and drug-induced lymphocyte stimulation test (DLST) against LCIG showed positive. Exchange of LCIG to LD drugs and intravenous methylprednisolone administration improved the symptoms and thrombocytopenia. CD induced type II and type IV allergy were suspected. This case offers a caution that physicians should be aware of drug allergy in cases of which unexpected symptoms occurred in altering one LD compound to another.
Topics: Male; Humans; Aged; Carbidopa; Levodopa; Antiparkinson Agents; Hyperthermia; Lymphocyte Activation; Parkinson Disease; Hypersensitivity; Drug Combinations; Syndrome; Thrombocytopenia
PubMed: 38092414
DOI: 10.5692/clinicalneurol.cn-001883 -
MSphere Jan 2024Parkinson's disease (PD) is characterized by motor symptoms and a loss of dopaminergic neurons, as well as a variety of non-motor symptoms, including constipation,...
Parkinson's disease (PD) is characterized by motor symptoms and a loss of dopaminergic neurons, as well as a variety of non-motor symptoms, including constipation, depression, and anxiety. Recently, evidence has also accumulated for a link between gut microbiota and PD. Most PD patients are on dopamine replacement therapy, primarily a combination of L-DOPA and carbidopa; however, the effect of these medications on the microbiota and non-motor symptoms in PD is still unclear. In this study, we explored the effects of chronic oral treatment with L-DOPA plus carbidopa (LDCD) on the gut microbiota and non-motor symptoms in males of a transgenic mouse model of PD (dbl-PAC-Tg();-/-). To further test whether the effects of these PD medications were mediated by the gut microbiota, oral antibiotic treatment (Abx; vancomycin and neomycin) was included both with and without concurrent LDCD treatment. Post-treatment, the gastrointestinal, motor, and behavioral phenotypes were profiled, and fecal, ileal, and jejunal samples were analyzed for gut microbiota composition by 16S sequencing. LDCD treatment was found to improve symptoms of constipation and depression in this model, concurrent with increases in abundance in the ileum. Abx treatment worsened the symptoms of constipation, possibly through decreased levels of short-chain fatty acids and disrupted gut barrier function. LDCD + Abx treatment showed an interaction effect on behavioral symptoms that was also associated with ileal levels. This study demonstrates that, in a mouse model, PD medications and antibiotics affect PD-related non-motor symptoms potentially the gut microbiota.IMPORTANCEThe motor symptoms of Parkinson's disease (PD) are caused by a loss of dopamine-producing neurons and are commonly treated with dopamine replacement therapy (L-DOPA plus carbidopa). PD has also been associated with altered gut microbiota composition. However, the effects of these PD medications on PD-related non-motor symptoms and the gut microbiota have not been well characterized. This study uses a transgenic mouse model of PD to help resolve medication-induced microbiota alterations from those that are potentially disease relevant within a PD context, and explores how long-term treatment may interact with the gut microbiota to impact non-motor symptoms.
Topics: Humans; Male; Mice; Animals; Parkinson Disease; Levodopa; Carbidopa; Mice, Transgenic; Dopamine; Anti-Bacterial Agents; Gastrointestinal Microbiome; Constipation
PubMed: 38078745
DOI: 10.1128/msphere.00379-23 -
Analytica Chimica Acta Jan 2024A sensitive method for the detection of β-glucuronidase was established using functionalized carbon dots (β-CD-SiCDs) as fluorescent probes. The β-CD-SiCDs were found...
A sensitive method for the detection of β-glucuronidase was established using functionalized carbon dots (β-CD-SiCDs) as fluorescent probes. The β-CD-SiCDs were found to be obtained through in situ autopolymerization by mixing the solutions of methyldopa, mono-6-ethylenediamine-β-cyclodextrin and N-(β-aminoethyl)-γ-aminopropyltrimethoxysilane at room temperature. The method has the characteristics of low energy consumption, simple and rapid. β-CD-SiCDs exhibited green fluorescence at 515 nm emission with a quantum yield of 7.9 %. 4-nitrophenyl-β-D-glucuronide was introduced as a substrate for β-glucuronidase to generate p-nitrophenol. Subsequently, p-nitrophenol self-assembled with β-CD-SiCDs through host-guest recognition to form a stable inclusion complex, resulting in the fluorescence quenching of β-CD-SiCDs. The linear range of β-CD-SiCDs for detecting β-glucuronidase activity was 0.5-60 U L with a detection limit of 0.14 U L. For on-site detection, gel reagents were prepared by a simple method and the images were visualized and quantified by taking advantage of smartphones, avoiding the use of large instrumentation. The constructed fluorescence sensing platform has the benefits of easy operation and time saving, and has been successfully used for the detection of β-glucuronidase activity in serum and cell imaging.
Topics: Glucuronidase; Cyclodextrins; Quantum Dots; Carbon; Fluorescent Dyes
PubMed: 38057046
DOI: 10.1016/j.aca.2023.341996 -
Sensors (Basel, Switzerland) Nov 2023The quality of life of patients affected by Parkinson's disease is improved by medications containing levodopa and carbidopa, restoring the dopamine concentration in the...
The quality of life of patients affected by Parkinson's disease is improved by medications containing levodopa and carbidopa, restoring the dopamine concentration in the brain. Accordingly, the affordable quality control of such pharmaceuticals is very important. Here is reported the simple and inexpensive colorimetric quantification of carbidopa in anti-Parkinson drugs by the selective condensation reaction between the hydrazine group from carbidopa and the formyl functional group of selected aldehydes in acidified hydroalcoholic solution. An optical assay was developed by using indole-3-carbaldehyde (I3A) giving a yellow aldazine in EtOH:HO 1:1 (λ~415 nm) at 70 °C for 4 h, as confirmed by LC-MS analysis. A filter-based plate reader was used for colorimetric data acquisition, providing superior results in terms of analytical performances for I3A, with a sensitivity ~50 L g and LOD ~0.1 mg L in comparison to a previous study based on vanillin, giving, for the same figures of merit values, about 13 L g and 0.2-0.3 mg L, respectively. The calibration curves for the standard solution and drugs were almost superimposable, therefore excluding interference from the excipients and additives, with very good reproducibility (RSD% 2-4%) within the linear dynamic range (10 mg L-50 mg L).
Topics: Humans; Carbidopa; Quality of Life; Reproducibility of Results; Colorimetry; Antiparkinson Agents; Levodopa
PubMed: 38005530
DOI: 10.3390/s23229142 -
CNS Neuroscience & Therapeutics Apr 2024This study aimed to systematically compare the effectiveness, safety, and costs of different anti-Parkinson drugs (APDs).
AIMS
This study aimed to systematically compare the effectiveness, safety, and costs of different anti-Parkinson drugs (APDs).
METHODS
This is a multi-center study that retrospectively analyzed the data of 8420 outpatients with PD from 2014 to 2019 across 30 tertiary hospitals in China. The effectiveness was evaluated by changes in total dosages of APDs, normalized by levodopa equivalent dose (LED) and presented as ΔLEDs; levodopa equivalent dose cost (LEDc) represented the daily cost of APDs; and newly added diagnostics were represented as APDs-related adverse events.
RESULTS
A total of 384 patients with eligible medical records for three consecutive years were enrolled. Patients treated with carbidopa/levodopa or levodopa/benserazide had significantly lower mean ΔLEDs than other groups (p < 0.01), followed by pramipexole and selegiline. The piribedil group had the highest ΔLEDs, with mean differences of 112.56-355.04 mg compared to other groups (p < 0.01). Meanwhile, LEDc in the levodopa/benserazide, carbidopa/levodopa, and piribedil groups were significantly lower than those in pramipexole or selegiline groups ($0.088-0.135/day for levodopa/benserazide; $0.070-0.126/day for carbidopa/levodopa; $0.112-0.138/day for piribedil; $0.290-0.332/day for pramipexole; $0.229-0.544/day for selegiline; p < 0.01). Patients with piribedil had more adverse events, with an incidence rate of 35.7%, followed by levodopa/benserazide (25.6%), selegiline (23.5%), carbidopa/levodopa (23.3%), and pramipexole (16.4%). Pramipexole showed a lower incidence rate of adverse events than piribedil, including neuropsychiatric symptoms (p = 0.006), headache/dizziness (p = 0.016), and gastrointestinal symptoms (p = 0.031).
CONCLUSIONS
Carbidopa/levodopa or levodopa/benserazide might exhibit better clinical improvement with less medical cost, while piribedil presented less clinical improvement but a higher risk of headache/dizziness, gastrointestinal, and neuropsychiatric symptoms.
Topics: Humans; Levodopa; Carbidopa; Benserazide; Retrospective Studies; Pramipexole; Parkinson Disease; Piribedil; Selegiline; Dizziness; Antiparkinson Agents; Headache
PubMed: 37983933
DOI: 10.1111/cns.14531 -
Journal of Molecular Recognition : JMR Mar 2024Enzymes are usually stereospecific against chiral substrates, which is commonly accepted for the amine oxidase family of enzymes as well. However, the FsqB (fumisoquin...
Enzymes are usually stereospecific against chiral substrates, which is commonly accepted for the amine oxidase family of enzymes as well. However, the FsqB (fumisoquin biosynthesis gene B) enzyme that belongs to the family of sarcosine oxidase and oxidizes L-N-methyl-amino acids, shows surprising activity for both enantiomers of N-methyl-dopa. The aim of this study is to understand the mechanism behind this behavior. Primary docking experiments showed that tyrosine and aspartate residues (121 and 315 respectively) are located on the ceiling of the active site of FsqB and may play a role in fixing the N-methyl-dopa via its catechol moiety and allowing both stereoisomers of this substrate to be in close proximity of the N5 atom of the isoalloxazine ring of the cofactor. Three experimental approaches were used to prove this hypothesis which are: (1) studying the oxidative ability of the variants Y121F and D315A on N-methyl-dopa substrates in comparison with N-methyl-tyrosine substrates; (2) studying the FsqB WT and variants catalyzed biotransformation via high-performance liquid chromatography (HPLC); (3) molecular dynamics simulations to characterize the underlying mechanisms of the molecular recognition. First, we found that the chemical characteristics of the catechol moiety of N-methyl-dopa are important to explain the differences between N-methyl-dopa and N-methyl-tyrosine. Furthermore, we found that Y121 and D315 are specific in FsqB and not found in the model enzyme sarcosine oxidase. The on-bench and theoretical mutagenesis studies show that Y121 residue has a major role in fixing the N-methyl-dopa substrates close to the N5 atom of the isoalloxazine ring of the cofactor. Simultaneously, D315 has a supportive role in this mechanism. Jointly, the experimental and theoretical approaches help to solve the riddle of FsqB amine oxidase substrate specificity.
Topics: Sarcosine Oxidase; Aspergillus fumigatus; Stereoisomerism; Fungal Proteins; Azoles; Drug Resistance, Fungal; Tyrosine; Methyldopa; Kinetics
PubMed: 37968575
DOI: 10.1002/jmr.3068