ABSTRACT
Atomoxetine is a psychostimulant drug used for the treatment of attention-deficit/hyperactivity disorder (ADHD) symptoms in people with autism. Herein, eco-friendly fluorescent carbon quantum dots (CQDs) were synthesized using black-eyed pea beans and characterized for the purpose of quantifying atomoxetine in pharmaceutical capsules and human plasma. The selectivity of these CQDs towards atomoxetine was improved by functionalizing their surface with an atomoxetine-tetraphenylborate ion complex. The quantification of atomoxetine is based on measuring the fluorescence quenching of the functionalized CQDs in response to varying concentrations of atomoxetine. The Stern-Volmer plot was employed to investigate the mechanism through which atomoxetine quenches the fluorescence intensity of the CQDs. The outcomes indicated a dynamic quenching mechanism. The applied method was optimized and validated in compliance with ICH requirements, resulting in excellent linearity across the concentration range of 50-800 ng/mL. The developed method was successfully used to quantify atomoxetine in pharmaceutical dosage form and human plasma with acceptable accuracy and precision outcomes. In addition, the method was applied for clinical pharmacokinetic study of atomoxetine in the plasma of children diagnosed with both autism and ADHD. Atomoxetine was rapidly absorbed after a single oral dose of 10 mg, reaching maximum concentration within two hours and having a half-life (t1/2) of 3.11 h. Moreover, the method demonstrates a notable degree of eco-friendliness, as evidenced by two greenness evaluation metrics; Green Analytical Procedure Index (GAPI) and Analytical GREEnness (AGREE).
Subject(s)
Atomoxetine Hydrochloride , Fluorescent Dyes , Quantum Dots , Spectrometry, Fluorescence , Atomoxetine Hydrochloride/pharmacokinetics , Atomoxetine Hydrochloride/blood , Humans , Quantum Dots/chemistry , Fluorescent Dyes/chemistry , Fluorescent Dyes/pharmacokinetics , Spectrometry, Fluorescence/methods , Attention Deficit Disorder with Hyperactivity/drug therapy , Attention Deficit Disorder with Hyperactivity/blood , Child , Male , Autistic Disorder/drug therapy , Reproducibility of Results , Limit of DetectionABSTRACT
AIM/PURPOSE OF STUDY: It is estimated that around 15 million babies are born prematurely every year and approximately one million children die each year due to complications of preterm birth (PTB). Many survivors face a lifetime of disability, including learning disabilities and visual and hearing problems. The current study aimed to characterize Lactobacillus species isolated from vaginal swabs and determine their antibiotic susceptibility. MATERIAL AND METHODS: 40 term and 20 preterm samples were processed by culturing on MRS agar and initial identification was carried out using sugar fermentation reactions and 16S rRNA PCR. Moreover, Lactobacillus species from preterm and term cases using paired samples, i.e. vaginal swabs and placenta tissues from 8 preterm delivering mothers were further recruited for metagenomics study to possibly detect uncultured Lactobacillus species known to cause PTB. RESULTS: 40% samples from preterm delivering mothers lack any Lactobacillus species whereas in contrast vaginal microflora of all term delivering mothers carry one or more species of Lactobacillus. L. crispatus (46% in term group and 25% preterm group) was found to be the most abundant group followed by L. jensenii (25% in both groups) and L. gasseri (19% in term group and 10% in preterm group). The antimicrobial susceptibility profile suggests that preterm isolates were least resistant to linezolid (20% resistance rate) and trimethoprim/sulfamethoxazole (22% resistant rate) compared to term isolates (>60% for each drug group). Furthermore, the metagenomics data for paired samples (8 VS and 8 PT) from extreme PTB suggested that Lactobacillus Iners was the main difference between term and preterm deliveries. Moreover, overall lack of lactobacillus species or presence of rogue Lactobacillus species such as L. iners and L. vaginilis is associated with PTB. ERIC-PCR analysis using Lactobacillus crispatus revealed that all the pre-term samples are closely related and fall in same cluster while all the term samples fall in different cluster. CONCLUSION: The study not only provides a baseline data of distinct signatures of associated lactobacillus species with the PTB which may be further transmitted to new born infants but also developing further therapeutic interventions to better manage the PTB.
Subject(s)
Premature Birth , Child , Female , Humans , Infant, Newborn , Lactobacillus/genetics , Pakistan , Pregnancy , Premature Birth/epidemiology , RNA, Ribosomal, 16S/genetics , VaginaABSTRACT
Biosensors based on microbial cells have been developed to monitor environmental pollutants. These biosensors serve as inexpensive and convenient alternatives to the conventional lab based instrumental analysis of environmental pollutants. Small monomeric naturally occurring fluorescent proteins (fp) can be exploited by converting them as small biosensing devices for biomedical and environmental applications. Moreover, they can withstand exposure to denaturants, high temperature, and a wide pH range variation. The current study employs newly identified novel fluorescent protein HriGFP from Hydnophora rigida to detect environmental contaminants like heavy metals and organo-phosphorous (pesticide) compounds such as methyl parathion. The HriGFP was initially tested or its expression in bacterial systems (Gram positive and Gram negative) and later on for its biosensing capability in E coli (BL21DE3) for detection of heavy metals and methyl parathion was evaluated. Our results indicated the discrete and stable expression of HriGFP and a profound fluorescent quenching were observed in the presence of heavy metals (Hg, Cu, As) and methyl parathion. Structural analysis revealed heavy metal ions binding to HriGFP via amino acid residues. In-silico-analysis further revealed strong interaction via hydrogen bonds between methyl parathion phosphate oxygen atoms and the amino group of Arg119 of HriGFP. This study implies that HriGFP can act as a biosensor for detecting harmful carcinogenic pesticide like methyl parathion in water resources in the vicinity of heavily pesticide impregnated agricultural lands and heavy metal contaminated water bodies around industrial areas.
Subject(s)
Anthozoa/metabolism , Biosensing Techniques/methods , Luminescent Proteins/genetics , Luminescent Proteins/metabolism , Animals , Anthozoa/genetics , Bacillus megaterium/genetics , Bacillus megaterium/growth & development , Binding Sites , Escherichia coli/genetics , Escherichia coli/growth & development , Hydrogen Bonding , Luminescent Proteins/chemistry , Metals, Heavy/analysis , Metals, Heavy/chemistry , Models, Molecular , Organophosphorus Compounds/analysis , Organophosphorus Compounds/chemistryABSTRACT
Primary cardiac tumours are an uncommon cause of intra-cardiac mass. Most common intra-cardiac mass is thrombus followed by infiltrating secondary cardiac tumours. These secondary tumours are more likely to invade the cardiac chambers when they arise from close vicinity of the heart. We report an unusual case of mediastinal lymphoma in a 55 years old lady presenting as an intra-cardiac mass in the right atrium, which was detected by a transthoracic echocardiogram and later confirmed with more specific modalities.