Search Results (181)

Narcotics trafficking is a fundamental part of organized crime, posing significant and evolving challenges for forensic investigations. Addressing these challenges requires rapid, precise, and scientifically validated analytical methods for reliable identification of illicit substances. Over the past five years, forensic drug testing has advanced considerably, improving detection of traditional drugs—such as tetrahydrocannabinol, cocaine, heroin, amphetamine-type stimulants, and lysergic acid diethylamide—as well as emerging new psychoactive substances (NPS), including synthetic cannabinoids (e.g., 5F-MDMB-PICA), cathinones (e.g., α-PVP), potent opioids (e.g., carfentanil), designer psychedelics (e.g., 25I-NBOMe), benzodiazepines (e.g., flualprazolam), and dissociatives (e.g., 3-HO-PCP). Current technologies include colorimetric assays, ambient ionization mass spectrometry, and chromatographic methods coupled with various detectors, all enhancing accuracy and precision. Vibrational spectroscopy techniques, like Raman and Fourier transform infrared spectroscopy, have become essential for non-destructive identification. Additionally, new sensors with disposable electrodes and miniaturized transducers allow ultrasensitive on-site detection of drugs and metabolites. Advanced chemometric algorithms extract maximum information from complex data, enabling faster and more reliable identifications. An important emerging trend is the adoption of green analytical methods—including direct analysis, solvent-free extraction, miniaturized instruments, and eco-friendly chromatographic processes—that reduce environmental impact without sacrificing performance. This review provides a comprehensive overview of innovations over the last five years in forensic drug analysis based on the ScienceDirect database and highlights technological trends shaping the future of forensic toxicology. Full article

Early diagnosis is critical for the effective management of neurodegenerative disorders, and retinal alterations have emerged as promising early biomarkers due to the retina’s close developmental and functional link to the brain. The zebrafish (Danio rerio), with its rapid development, transparent embryos, and evolutionarily conserved visual system, represents a powerful and versatile model for studying retinal degeneration. This review discusses a range of behavioral assays—including visual adaptation, motion detection, and color discrimination—that are employed to evaluate retinal function in zebrafish. These methods enable the detection of subtle visual deficits that may precede overt anatomical damage, providing a non-invasive, efficient strategy for early diagnosis and high-throughput drug screening. Importantly, these behavioral tests also serve as sensitive functional readouts to evaluate the efficacy of pharmacological treatments over time. Compared to traditional murine models, zebrafish offer advantages such as lower maintenance costs, faster development, optical transparency for live imaging, and ethical benefits due to reduced use of higher vertebrates. However, variability in experimental protocols highlights the need for standardization to ensure reliability and reproducibility. Full article

Background: Accurate and rapid diagnosis of drug-resistant tuberculosis is essential for initiating appropriate treatment and preventing the transmission of these strains. This study compares phenotypic and genotypic methods of drug susceptibility testing for Mycobacterium tuberculosis (M. tuberculosis). Methods: Resistance to first-line drugs, as well as resistance to second-line drugs (fluoroquinolones and aminoglycosides), was assessed using the Löwenstein–Jensen medium phenotypic method and the GenoType MTBDRplus genotypic method and analyzed. Results: The phenotypic resistance rate was 84.85% for INH (n = 56), 46.97% for RIF (n = 31), 48.48% for STR (n = 32), and 30.30% for EMB (n = 20). Of the MDR-TB isolates (n = 29), 41.37% were resistant to fluoroquinolones (n = 12) and 31.03% were resistant to both fluoroquinolones and injectable aminoglycosides, being classified as XDR-TB (n = 9). In addition, 22.73% of the MDR-TB isolates were resistant to all four first-line drugs (n = 15). The overall concordance between the line probe assay method and phenotypic testing was 94.74% for RIF and 95.16% for INH. Discordances were identified in three cases for RIF and two cases for INH, where isolates were reported as susceptible by GenoType MTBDRplus, but phenotypically resistant. Conclusions: Genotypic testing using GenoType MTBDRplus provides rapid and accurate results, but some cases of phenotypic resistance are not detected by this method. The results highlight the importance of using combined phenotypic and genotypic methods for accurate diagnosis of MDR-TB, as well as the need to integrate genomic sequencing to improve diagnostic accuracy. Full article

Background/Objectives: Over the past few decades, controlled substance abuse in drug-facilitated sexual assaults (DFSAs) has significantly increased worldwide, leading to an urgency to develop rapid and selective drug detection methods for field use (i.e., on-spot detection). Currently, techniques for detecting DFSA drug-associated samples are laborious and require skilled personnel to analyze/interpret the results. Moreover, most DFSA-associated drugs have a short half-life, making them more challenging to detect promptly. For instance, the timely detection of gamma-hydroxybutyrate (GHB) has been of ultimate concern for decades due to its fast elimination from the body. This study describes the development of a fluorescent ionic liquid nanosensor that can be used to rapidly detect GHB in the field. Methods: Trihexyltetradecylphosphonium fluorescein (THP₂FL) ionic liquid was synthesized and evaluated for its potential application in detecting GHB. THP₂FL nanoparticles in deionized water were synthesized with a size of 199 nm by a reprecipitation method. Results: The addition of GHB to THP₂FL nanoparticles resulted in up to a 60% increase in fluorescence intensity and a 79% increase in absorbance. These results suggest potential applications for using the fluorescent THP₂FL nanoparticles to detect GHB. The sensor’s selectivity was tested on compounds structurally similar to GHB, and the results showed that 1,4-butanediol (a precursor of GHB) is a potentially interfering species. Conclusion: This fluorescent technique allows for field deployable sensors, which would benefit screening GHB onsite. Full article

The integration of metal–organic frameworks (MOFs) with microfluidic technologies has opened new frontiers in biomedical diagnostics and therapeutics. Microfluidic chips offer precise fluid control, low reagent use, and high-throughput capabilities features further enhanced by MOFs’ ample surface area, adjustable porosity, and catalytic activity. Together, they form powerful lab-on-a-chip platforms for sensitive biosensing, drug delivery, tissue engineering, and microbial detection. This review highlights recent advances in MOF-based microfluidic systems, focusing on material innovations, fabrication methods, and diagnostic applications. Particular emphasis is placed on MOF nanozymes, which enhance biochemical reactions for multiplexed testing and rapid pathogen identification. Challenges such as stability, biocompatibility, and manufacturing scalability are addressed, along with emerging trends like responsive MOFs, AI-assisted design, and clinical translation strategies. By bridging MOF chemistry and microfluidic engineering, these systems hold great promise for next-generation biomedical technologies. Full article

Tuberculosis (TB), primarily caused by Mycobacterium tuberculosis complex (MTBC), remains a global health challenge and can lead to severe pulmonary and extrapulmonary complications. Multidrug-resistant TB (MDR-TB) poses additional challenges, requiring advanced diagnostic and treatment strategies. This study evaluates the BD MAX MDR-TB molecular test for a rapid diagnosis of MDR-TB, detecting resistance to rifampicin (RIF) and isoniazid (INH). The BD MAX MDR-TB test, utilizing real-time PCR, was used to analyze specimens collected from TB-suspected patients, identifying MTB DNA and mutations associated with rifampicin and isoniazid resistance. Results were compared with traditional drug susceptibility testing, and 79 out of 638 samples tested were positive for MTB DNA, with 65 showing a sufficient amount of genetic material for resistance gene identification. The BD MAX test showed a 100% correlation with phenotypic rifampicin resistance, though discrepancies were noted for isoniazid resistance, with a 93% concordance. The BD MAX MDR-TB test is an effective tool for a rapid diagnosis of MDR-TB, especially for rifampicin resistance. However, it may not detect certain mutations related to isoniazid resistance. Complementary tests like Xpert MTB/XDR or whole-genome sequencing could improve diagnostic accuracy and support more effective TB control strategies. Full article

Background/Objectives: Voriconazole (VRCZ) use requires accurate monitoring to avoid suboptimal drug levels and adverse effects. In addition, the appearance of resistant fungal strains is a problem that needs attention. Blood concentration measurement is the monitoring technique of choice; however, it is slow, limiting its clinical application. This study aimed to evaluate the clinical utility of rapid therapeutic drug monitoring (TDM) for VRCZ using high-performance liquid chromatography with ultraviolet detection (HPLC-UV) compared to conventional outsourced liquid chromatography–tandem mass spectrometry (LC-MS/MS) testing in outpatient care. Methods: VRCZ blood concentrations were measured using HPLC-UV and LC-MS/MS. Reporting times, accuracy, and clinical outcomes were assessed for outpatients receiving VRCZ treatment. Safety was monitored for renal, hepatic, and visual toxicities. Results: HPLC-UV significantly reduced reporting times (0.433 h vs. 74.3 h, p < 0.001), and Deming’s regression analyses showed a strong correlation with LC-MS/MS results (Pearson’s r = 0.988). Bland–Altman analysis showed an average difference of 0.025 μg/mL between HPLC-UV and LC-MS/MS. Prospective monitoring of three outpatients revealed no adverse events, enabling safe and effective VRCZ dosing. Conclusions: Rapid VRCZ TDM using HPLC-UV is a cost-effective and feasible approach for outpatient care, significantly improving reporting times and patient safety. Further studies and cross-facility collaboration are needed to expand its application. Full article

Background: Extrapulmonary tuberculosis (TB) presents significant diagnostic challenges, particularly in the context of multidrug-resistant (MDR) strains. This study assessed the utility of the WHO-recommended rapid molecular assays, originally validated for pulmonary TB, in diagnosing extrapulmonary TB and detecting the MDR Mycobacterium tuberculosis complex (MTBC). Materials and Methods: A total of 6274 clinical samples, including 4891 pulmonary and 1383 extrapulmonary samples, were analyzed between 2019 and 2022 using the BD MAX™ MDR-TB assay (BD MAX), the Xpert^® MTB/RIF assay (Xpert MTB/RIF), the Xpert^® MTB/XDR assay (Xpert MTB/XDR), FluoroType MTB, and phenotypic drug susceptibility testing (DST). Results: MTBC was detected in 426 samples using BD MAX (376 pulmonary and 50 extrapulmonary), of which 277 were culture-confirmed. Phenotypic testing confirmed 299 positive cultures on Löwenstein–Jensen (LJ) medium and 347 in BD BACTEC™ MGIT™ (BACTEC MGIT) mycobacterial growth indicator tube (BBL) liquid culture. BD MAX showed high sensitivity and specificity for extrapulmonary TB detection (93.1% and 98.4%, respectively). Resistance to isoniazid or rifampicin was identified in 11% of MTBC-positive cases, whereas 3.69% were confirmed as MDR-TB. The molecular assays effectively detected resistance-associated mutations (katG, inhA, and rpoB), with high concordance to phenotypic tests (DST) (κ = 0.69–0.89). Conclusions: This study demonstrates that molecular assays, although validated for pulmonary TB, are also reliable for extrapulmonary TB detection and drug resistance profiling. Their rapid turnaround and robust accuracy support broader implementation in routine diagnostics, especially for challenging extrapulmonary specimens where early detection is critical for targeted therapy. Full article

Synthetic cannabinoids (SCs), a class of widely abused new psychoactive substances, are characterized by their structural diversity and rapid evolution. Structure–affinity relationships are crucial for predicting pharmacological effects and potential toxicity. Traditional methods for affinity testing are often complex and less applicable to newly modified compounds. In contrast, Surface Plasmon Resonance (SPR) is a sensitive and label-free technology that detects molecular interactions by measuring refractive index changes on a metallic surface with the advantages of high sensitivity, low sample consumption, and high-throughput capability. In this study, we used SPR to determine the receptor affinity constants of 10 SCs, including some first-reported substances, and analyzed their structure–affinity relationships to validate the method’s reliability. The results showed that (1) indazole-based SCs exhibited stronger CB1 receptor affinity compared to their indole counterparts, (2) the head structure of p-fluorophenyl enhanced affinity relative to 5-fluoropentyl, (3) and the affinity rankings obtained from SPR experiments were consistent with those derived from traditional methods. These results collectively demonstrate the reliability and effectiveness of SPR in assessing CB1 receptor affinity and differentiating affinity differences among structurally similar analogs, with promising application prospects in drug research, particularly in the development and screening of therapeutic agents targeting cannabinoid receptors. Full article

Background: Bacterial endophthalmitis is a rare but serious complication of intravitreal injections (IVIs). Prefilled syringes have been introduced to reduce contamination risk during drug preparation. However, whether they lower the incidence of bacterial endophthalmitis compared to vials remains unclear. Methods: This retrospective cohort study analyzed aflibercept IVIs performed at 17 clinical centers in Japan between 2015 and 2022. Patients aged ≥20 years who received aflibercept IVIs (vial or prefilled syringe) for age-related macular degeneration, diabetic macular edema, retinal vein occlusion, or myopic choroidal neovascularization were included. Bacterial endophthalmitis was diagnosed based on clinical signs (e.g., rapid vision loss, pain, hypopyon, vitreous opacity). Incidence rates were compared using Fisher’s exact test. Results: Among 152,039 injections (43,684 prefilled syringes; 108,355 vials), 12 cases of bacterial endophthalmitis were identified (0.0046% vs. 0.0092%, p = 0.53). Poor visual outcomes were associated with Enterococcus faecalis, Streptococcus spp., and diabetes. Conclusions: Although incidence was lower in the prefilled syringe group, the difference was not statistically significant. Detecting a significant difference requires a larger sample. Further studies are needed to confirm the potential benefits of prefilled syringes in reducing endophthalmitis risk. Full article

Background/Objectives: Ambroxol hydrochloride (AMB) is a promising chaperone for treating neurological manifestations in Gaucher disease type 3 (GD3). The Amsterdam University Medical Center planned to conduct an n-of-1 clinical trial using high-dose AMB (25 mg/kg/day). As an adequate commercial AMB formulation is unavailable for this high target dosage, we aimed to develop high-dose AMB capsules and assess the formulated capsule’s quality. Methods: AMB API was sourced and tested according to the requirements of the European Pharmacopoeia. Capsule formulations of 75 mg and 200 mg AMB were developed. Drug product specifications were set following international guidelines (ICH Q6A) and the European Pharmacopoeia. Analytical methods were developed and validated, and three validation batches of each capsule strength were produced and analyzed. Results: The contents and the Acceptance Values (AVs) of the initial AMB batches (both strengths) varied between 89.1% to 92.7% (specification: 90% to 110%) and 12.4 to 17.6 (specification ≤ 15.0), respectively, indicating non-uniform AMB distribution. Consequently, the production of 200 mg capsules was discontinued, and modifications were made to the 75 mg capsule formulation, followed by the production of three optimized 75 mg validation batches. These batches met the specified criteria, with an AMB content and AV values ranging from 93.9% to 96.5% and 12.4 to 14.9, respectively. Furthermore, rapid dissolution profiles were observed (>80% dissolution within 15 min). No degradation products or microbiological impurities were detected after production. Conclusions: The optimized formulation of 75 mg AMB capsules formulated within the hospital pharmacy setting resulted in qualitative and uniform capsules which can be used in clinical trials. Full article

Voltage-gated sodium (Nav) channels play a crucial role in initiating and propagating action potentials throughout the heart, muscles and nervous systems, making them targets for a number of drugs and toxins. While patch-clamp electrophysiology is considered the gold standard for measuring ion channel activity, its labor-intensive and time-consuming nature highlights the need for fast screening strategies to facilitate a preliminary selection of potential drugs or hazards. In this study, a high-throughput and cost-effective biosensing method was developed to rapidly identify specific agonists and inhibitors targeting the human Nav1.1 (hNav1.1) channel. It combines a red fluorescent dye sensitive to transmembrane potentials with CHO cells stably expressing the hNav1.1 α-subunit (hNav1.1-CHO). In the initial screening mode, the tested compounds were mixed with pre-equilibrated hNav1.1-CHO cells and dye to detect potential agonist effects via fluorescence enhancement. In cases where no fluorescence enhancement was observed, the addition of a known agonist veratridine allowed the indication of inhibitor candidates by fluorescence reduction, relative to the veratridine control without test compounds. Potential agonists or inhibitors identified in the initial screening were further evaluated by measuring concentration–response curves to determine EC₅₀/IC₅₀ values, providing semi-quantitative estimates of their binding strength to hNav1.1. This robust, high-throughput biosensing assay was validated through comparisons with the patch-clamp results and tested with 12 marine toxins, yielding consistent results. It holds promise as a low-cost, rapid, and long-term stable approach for drug discovery and non-target screening of neurotoxins. Full article

The accurate detection of Mycobacterium tuberculosis (MTB) is a pressing challenge in the precise prevention and control of tuberculosis. Currently, the efficiency and accuracy of drug resistance detection for MTB are low, and cross-contamination is common, making it inadequate for clinical needs. This study developed a rapid nucleic acid detection method for MTB based on scattering loop-mediated isothermal amplification (LAMP). Specific primers for the MTB-specific gene (Ag85B) were designed, and the LAMP reaction system was optimized using a self-developed scattering LAMP turbidimeter. Experimental results showed that the optimal reaction system included 1.5 µL of 100 mmol/L magnesium ions, 3.5 µL of 10 mmol/L dNTPs, 6 µL of 1.6 mol/L betaine, and a reaction temperature of 65 °C. The minimum detection limit was 12.40 ng/L, with the fastest detection time being approximately 10 min. The reaction exhibited good specificity, with no amplification bands for other pathogens. Twenty culture-positive samples and twenty culture-negative samples were tested in parallel; the accuracy of the positive group was 100%, the detection time was (24.9 ± 13 min), and there was no negative detection. This method features high detection efficiency, low cost, high accuracy, and effectively reduces cross-contamination, providing a new technology for the rapid clinical detection of MTB. Full article

Background: Point-of-care diagnostic tests are essential for confirming Onchocerca volvulus transmission in remote, resource-limited, onchocerciasis-endemic communities. In Maridi, South Sudan, we field-tested a novel “biplex A” rapid diagnostic test (RDT) developed by Drugs & Diagnostics for Tropical Diseases (DDTD), San Diego, California. Methods: In February 2023, children aged 3–9 years were recruited from study sites at different distances from the Maridi Dam, a known blackfly breeding site. O. volvulus antibodies were detected using the DDTD biplex A RDT, which detects antibodies to Ov16 and OvOC3261 at test line 1 and to Ov33.3 and OvOC10469 at test line 2, along with the commercially available Ov16 SD Bioline RDT. Both tests were performed on whole blood obtained via finger prick. The feasibility and acceptability of the DDTD biplex A RDT were assessed, and its results were compared with those of the Ov16 SD Bioline RDT. Results: A total of 239 children participated in the study. The anti-Ov16 seroprevalence detected by the Ov16 SD Bioline RDT was 30.2% (72/239), with the highest prevalence observed in children living closest to the Maridi Dam (p < 0.001). Testing with the DDTD biplex A RDT was determined to be feasible, acceptable, and easy to use in a field setting. The DDTD biplex A RDT test line 1 (anti-Ov16 and anti-OvOC3261) was positive in 35.1% (84/239) of children, while test line 2 (anti-Ov33.3 and anti-OvOC10469) was positive in 18.4% (44/239). Both lines were simultaneously visible in 15.5% (37/239). Conclusions: The DDTD biplex A RDT prototype was user-friendly and practical for field deployment. However, additional research is needed to evaluate its performance relative to the commercially available Ov16 SD Bioline RDT. The high anti-Ov16 seroprevalence that was observed underscores the ongoing O. volvulus transmission near the Maridi Dam. Strengthening the onchocerciasis elimination program in Maridi should be prioritized as a critical public health intervention. Full article

Occult HCV infection (OCI) is defined by the presence of HCV RNA in hepatocytes and/or peripheral blood mononuclear cells (PBMCs) without detectable HCV RNA or anti-HCV antibodies in plasma. OCI is underrecognized and may contribute to HCV transmission. This study estimated OCI prevalence and associated risk factors in adults from Mexico City. Methods: A retrospective cross-sectional study was conducted, analyzing 507 general population volunteers. Demographic data and potential risk factors were collected via questionnaire. Anti-HCV detection was performed using two techniques: immunochromatographic rapid test and chemiluminescent microparticle immunoassay (CMIA). Nested PCR was employed to detect HCV RNA in plasma and PBMCs. Positive samples were genotyped through sequencing and phylogenetic analysis of the Core/E1 region. Results: Of 507 participants, four were anti-HCV positive. HCV RNA was found in PBMCs of 27 individuals, while plasma samples tested negative, indicating a 5.3% OCI prevalence. OCI was significantly associated with blood donation (p = 0.015), drug use (p = 0.019), particularly cocaine (p = 0.001), and endoscopy (p = 0.043). Genotypes 1b, 1a, 2b, 3a, and 2j were detected in OCI cases. Conclusions: OCI prevalence in Mexico City’s general population is notable, with significant links to blood donation, cocaine use, and endoscopy. Enhanced diagnostic strategies are crucial to detect OCI and mitigate HCV transmission. Full article