Search Results (83)

White pine blister rust disease (WPBR), caused by Cronartium ribicola, ranks among the most destructive pathogens of five-needle pines. We developed a hydroxynaphthol blue (HNB)-based Loop-mediated isothermal amplification (LAMP) assay enabling rapid, visual detection of C. ribicola directly following DNA extraction. LAMP primers targeting the internal transcribed spacer (ITS) region were designed and validated through in silico comparison with related Cronartium species and in vitro testing against sympatric forest fungi. The optimized 25 μL reaction contained 8.0 mM Mg²⁺, 1.0 mM dNTPs, and an inner-to-outer primer ratio of 8:1, with amplification conducted at 62 °C for 40 min. Positive amplification produced a distinctive color transition from purple to sky blue, enabling visual interpretation without instrumentation. Under the tested conditions, the assay achieved a detection limit of 460 ± 3.2 fg/μL genomic DNA—a 10-fold improvement over conventional PCR in concentration-based sensitivity. Assay applicability was evaluated using 211 field-collected Pinus armandii samples sourced from China. Detection efficiency varied significantly across tissue types. Symptomatic bark exhibited a substantially higher positive detection rate (68.97%, 95% CI: 49.2–84.7%) compared to needles from symptomatic trees (18.75%, 95% CI: 4.1–45.7%). Among asymptomatic samples, 3.75% of bark samples tested positive for C. ribicola DNA, whereas all needle samples were negative. Geographically, positive detections clustered at several discrete sampling sites in southwestern China, predominantly at elevated elevations. The established LAMP-HNB assay provides a rapid, visually interpretable diagnostic tool for early detection and quarantine monitoring of WPBR following DNA extraction. Beyond its practical utility, this assay establishes valuable baseline data for targeted disease surveillance in the context of evolving climate conditions. Full article

Quality by design (QbD) is a systematic approach focused on achieving consistent, predictable quality based on predefined objectives. Unlike traditional methods, QbD prioritizes risk assessment and management, which significantly enhances the robustness of the analytical method. In this study, we initiated factor screening using a three-factor, two-level design to evaluate three independent variables: flow rate, pH, and mobile phase composition. To further investigate the interaction of these variables, we employed Central Composite Design (CCD). This allows us to apply response surface methodology to the Critical Analytical Attributes (CAAs), specifically retention time, peak area, and symmetry factor, by conforming to the method’s robustness. The combination of pregabalin and duloxetine hydrochloride (HCl) dosage form was determined using a straightforward, exact, specific, and accurate reverse-phase HPLC approach. The results showed retention times of 3.265 min and 4.318 min for duloxetine HCl and pregabalin, respectively. Pregabalin demonstrated linearity from 100 to 200 μg/mL (R² = 0.998), whilst duloxetine HCl demonstrated linearity between 20 and 120 μg/mL (R² = 0.997). Lower LOD values of 0.925 µg/mL and 0.853 μg/mL and LOQ values of 2.809 μg/mL and 2.587 μg/mL of pregabalin and duloxetine HCl, respectively, suggest good sensitivity for the technique. The drug content of the commercial formulation may thus be determined using the recommended method. This technique can be used for standard quality control studies to simultaneously estimate pregabalin and duloxetine HCl. The novelty of the present studies lies in the development of a robust RP-HPLC method for simultaneous estimation of pregabalin and duloxetine HCl using a systematic AQbD approach, enhancing robustness, reproducibility, and reliability, making it highly suitable for routine quality control and regulatory applications. Full article

Neurodegenerative conditions such as Alzheimer’s disease, Parkinson’s disease, Huntington’s disease, and amyotrophic lateral sclerosis (ALS) are devastating disorders with the gradual loss of neurons and cognitive or motor disability. This is a review article that develops the crucial functions of autophagy and proteostasis within the scope of the neurodegenerative disease. Autophagy is a very well-conserved cell process that poses a quality control checkpoint that allows for the degradation and recycling of damaged organelles and misfolded proteins. Chaperones, the ubiquitin–proteasome complexes, and endoplasmic reticulum-associated degradation (ERAD) are also referred to as proteostasis, which plays a key role in ensuring the correct protein folding properties and the prevention of toxic protein accumulation. This article offers a detailed discussion of the relationship between autophagy and proteostasis, illustrating the mechanisms of mutual cooperation of these processes, ensuring cellular homeostasis and inhibiting the formation of pathogenic protein aggregates. In addition, this review includes experimental data during preclinical studies and clinical trials and expounds on the therapeutic opportunities that show the potential of targeting autophagy and proteostasis to counter neurodegenerative disorders. Although research progress creates potential for new indicators, the issues and difficulties relating to the effects of regulating such complex cellular pathways are also discussed in the article. Finally, the review can add to the research of neurodegenerative disease mechanisms of autophagy and proteostasis as well as provide insights about the future of treatment and its implementation. Full article

Objectives: This in vitro study aimed to evaluate the effects of three framework orientation (FO) positions on an SLM building platform (Horizontal [H], Diagonal-45° [D45°], Diagonal-60° [D60°]) and two designs (with [B] or without [NB] stabilizing bars) on the fitting accuracy of digitally fabricated Co-Cr RPD frameworks. Materials and Methods: A custom RPD framework CAD was performed on a 3D-printed resin-model of an edentulous maxilla with three geometric tooth forms. A Co-Cr alloy was processed via SLM processing into 24 framework specimens, divided into three FO groups (n = 8: H, D45°, D60°) and two subgroups each (n = 4: B, NB). Qualitative/quantitative fit-evaluation was assessed using virtual framework-to-model seating and a custom digital protocol with GOM Inspect software (2018-Hotfix5, Rev.115656). Mean fitting distances were calculated from 220 equidistant points per specimen. Statistical comparisons were performed using ANOVA-on-ranks, Kruskal–Wallis multiple comparisons, and Bonferroni adjustment. Results: FO Sub-Group medians (Q1, Q3: 25% and 75% Quartiles) (mm) were: H/NB 0.150 (0.140, 0.164), H/B: 0.136 (0.121, 0.152), D45°/NB: 0.230 (0.219, 0.241), D45°/B: 0.144 (0.137, 0.154), D60°/NB:0.238 (0.232, 0.247), D60°/B: 0.171 (0.166,0.176). Pairwise comparisons indicated the following statistically significant (p < 0.05) FO Sub-Group differences: H/B-D45°/NB, H/B-D60°/NB, D45°/B-D45°/NB, D45°/B-D60°/NB, H/NB-D45°/NB, H/NB- D60°/NB. Conclusions: Horizontal orientation improved RPD fit accuracy regardless of bar presence. D45° accuracy is enhanced by stabilizing bars, while D60° accuracy is unaffected by bar addition. Full article

The aadA1 gene, which confers resistance to streptomycin, is typically located within class Ⅰ integrons. This genetic context enables its dissemination among diverse Gram-negative bacteria, such as Salmonella, and facilitates its potential transfer to humans through the food chain or into agricultural environments via manure. Hence, the detection of aadA1 genes is crucial for surveillance, understanding transmission dynamics, and informing strategies to mitigate the spread of resistant bacteria. Conventional aadA1 detection relies on time-consuming or equipment-intensive molecular assays like PCR or qPCR. In this study, we developed and optimized a closed-tube, hydroxynaphthol blue (HNB) -based loop-mediated isothermal amplification (LAMP) assay to detect Salmonella aadA1 gene and performed evaluation and validation against conventional PCR. The LAMP assay demonstrated high specificity and sensitivity, with a detection limit of 1 pg (190 copies) of genomic DNA per reaction, which is tenfold higher than that of conventional PCR. In parallel testing of 40 Salmonella DNA samples, the optimized LAMP assay achieved a detection rate of 100.0% for the aadA1 gene in streptomycin-resistant isolates, compared to 96.3% by conventional PCR. Among the streptomycin-susceptible isolates, the LAMP assay also showed a higher detection rate (38.5%) for the aadA1 gene, compared to 23.1% by conventional PCR. Consequently, the LAMP assay developed in this study for detecting the aadA1 gene offers a combination of simplicity, speed, visual readout, high specificity, and sensitivity, making it particularly suitable for rapid field detection in antimicrobial resistance surveillance programs. Full article

Quantitative determination of calcium content in milk and dairy products is an important analytical task due to the great importance of this element in the human diet. Calcium determination in milk and dairy products was performed using semi-automatic complexometric titration with a webcam as a color change detector. The color changes were registered directly in turbid dairy dispersions, creating a white background. The analytical signals tested were the color component (R, G, B) read from the titration beaker images and the calculated Hue parameter. For the calcein indicator, the optimal signals are Green and Hue. For the HNB indicator, the optimal signals are Red and Hue. The developed method of titration using an RGB camera detector is characterized by excellent linearity (R² = 0.9999) and accuracy. RSD values range from 0.3 to 2.9%. Recovery values range from 105 to 113%. Examples of calcium determination in commercial products include milk, fermented products, cream, and cottage cheese. Full article

Background and Clinical significance: Single-shot spinal anesthesia for intrapartum Cesarean section has recently been incriminated in carrying a high risk of high neuraxial block (HNB) occurrence in parturients receiving labor epidural analgesia. The so-called volume effect of the epidurally injected solution causes a contraction of the dural sack and unexpected HNB. Case presentation: We present two cases of HNB in parturients receiving epidural analgesia. The first case describes the 36-year-old patient G3P2, who was administered a repeated rescue analgesia single-shot spinal injection with low-dose local anesthetic (levobupivacaine, 3 mg) following non-functional combined spinal–epidural analgesia. The second case describes the 28-year-old parturient G1P0, who experienced HNB after single-shot spinal anesthesia with hyperbaric bupivacaine (7.5 mg) following labor epidural analgesia. Conclusions: Intrathecal administration of local anesthetic for the purpose of spinal analgesia or anesthesia in a parturient with epidural analgesia can cause unexpected HNB and could occur even at low doses of intrathecally administered medications. The interplay of numerous variables and circumstances in the specific case can result in the occurrence of HNB. We assume that in our first case, the volume effect and repeated dural puncture, and in the second case, the low height of the parturient coupled with the volume effect, played significant role in the occurrence of HNB. Full article

Niobium oxides are promising materials for catalytic applications due to their unique structural versatility and surface chemistry. Nb₂O₅ nanomaterials were synthesized via a solvothermal method at 150 °C using niobium oxalate as a precursor. A comprehensive characterization of the material was performed using electron microscopy, X-ray diffraction, and Raman spectroscopy. The as-prepared nanoparticles primarily crystallized in a mixture of the TT-Nb₂O₅ phase (TT from the German Tief-Tief, meaning “low-low”) and niobic acid, while subsequent thermal treatment at 900 and 1100 °C induced a phase transformation to T-Nb₂O₅ and H-Nb₂O₅, respectively (T from the German Tief, meaning “low”, and H from Hoch, meaning “high”). The as-prepared samples consist of micro-coils composed of interconnected nanometer-scale fibers, whereas the morphology changes into rods when they are treated at 1100 °C. The photocatalytic performance of the nanoparticles was evaluated by comparing the as-prepared and thermally treated samples. The as-prepared nanoparticles exhibited the highest photocatalytic activity under visible illumination, achieving 100% degradation after 180 min. More interestingly, the treatment of the as-prepared material with H₂O₂ modified the surface species formed on the Nb₂O₅, altering the photocatalytic behavior under various illumination conditions. This sample showed the highest photocatalytic activity under UV illumination, reaching 100% degradation after 75 min. On the other hand, the calcined samples are practically inactive, attributed to the loss of active catalytic sites during thermal treatment and phase transformation. Full article

Electroanalysis has emerged as a critical tool in the pharmaceutical industry, offering versatile and sensitive methods for drug analysis. This review explores the principles, techniques, and applications of electroanalysis in pharmaceuticals, emphasizing its role in drug development, quality assurance, pharmacokinetics, and environmental monitoring. Key electroanalytical methods, including voltammetry, potentiometry, and amperometry, are detailed along with their practical applications, such as detecting active pharmaceutical ingredients, monitoring drug metabolites, and ensuring product stability. Innovations in electrode materials and biosensors have enhanced their sensitivity and specificity, paving the way for advanced drug screening and therapeutic monitoring. Challenges like electrode fouling, selectivity issues, and regulatory constraints are discussed, along with strategies to overcome them. Future trends highlight the integration of nanotechnology, AI, and portable sensors to facilitate real-time analysis and personalized medicine. These advancements position electroanalysis as an indispensable component of modern pharmaceutical research and healthcare. Future perspectives emphasize the integration of nanotechnology and artificial intelligence (AI) to optimize experimental processes and data interpretation. This study also predicts the increased adoption of lab-on-a-chip systems and bioelectrochemical sensors to meet the growing demand for precision medicine and sustainable pharmaceutical practices. These advancements position electroanalysis as a cornerstone of pharmaceutical research, paving the way for more efficient drug development, improved patient outcomes and better environmental management. This comprehensive review underscores the transformative potential of electroanalysis in addressing the evolving challenges of the pharmaceutical industry and provides a foundation for future innovations. This review does not explicitly define the timeframe for the considered advancements. However, it discusses recent technological developments, including innovations in nanostructured electrodes, microfluidic integration, and AI-driven data analysis, indicating a focus on advancements primarily from the last few years, i.e., from 2020 to 2025. Full article

(1) Background: There are many cases of human disease caused by the hepatitis A virus contamination of aquatic products, so the development of the rapid detection of hepatitis A virus in aquatic products is crucial. (2) Methods: In this study, we developed three visual loop-mediated isothermal amplification methods for the rapid and intuitive detection of hepatitis A virus in aquatic products. New specific LAMP primers were designed for the HAV-specific VP1 protein shell. (1) HNB dye was added to the LAMP reaction system. After the reaction, the color of the reaction mixture changed from violet to sky blue, showing a positive result. (2) Cresol red dye was added to the LAMP reaction system, and a positive result was indicated by orange, while a negative result was indicated by purple. (3) By labeling FIP with biotin and LF with 6-FAM, the amplified product simultaneously contained biotin and 6-FAM, which bound to the anti-biotin antibody on the gold nanoparticles on the lateral flow dipstick (LFD). Subsequently, biotin was further combined with the anti-fam antibody on the T-line of the test strip to form a positive test result. (3) Results: The three visual LAMP methods were highly specific for HAV. The sensitivity of the visual assay was 2.59 × 10⁰ copies/μL. The positive detection ratio for 155 bivalve shellfish samples was 8.39%, which was the same as that for RT-qPCR. The three visual LAMP methods established in our work have better sensitivity than the international gold standard, and their operation is simple and requires less time. (4) Conclusions: The results can be obtained by eye color comparison and lateral flow dipsticks. Without the use of large-scale instrumentation, the sensitivity is the same as that of RT-qPCR. The test strips are lightweight, small in size, and easy to carry; they are suitable for emergency detection, on-site monitoring, field sampling, or remote farms and other non-laboratory environments for rapid identification. Full article

Background: Depression is one of the most serious and common health problems among the youth population and is responsible for the initiation of many diseases. As per the World Health Organization, 3.8% of the population suffers from mental depression, globally. The monoamine oxidase-A (MAO-A) enzyme is responsible for the degradation of neurotransmitters leading to lower levels of neurotransmitters. Methods: Chalcones (C1-C15) were synthesized by reacting substituted acetophenone with various benzaldehydes in a basic ethanolic solvent at 80 °C under microwave irradiation conditions. To compare the reaction time and product yield, a conventional method of synthesis of chalcones was also performed. The synthesized chalcones (C1-C15) were spectroscopically characterized and screened initially for inhibitory activities against MAO-A and MAO-B. The best active compounds were undertaken for IC₅₀ determination against MAO-A enzyme followed by the reversibility of inhibition analysis and the antioxidant assay. Moreover, in silico molecular docking and ADME pharmacokinetic investigations were accomplished. Results: Most of the compounds inhibited MAO-A, specifically, compounds C14 and C6 exhibited the highest inhibition at IC₅₀ values of 7.91 ± 0.08 μM and 8.45 ± 0.19 μM, respectively. Both these compounds exhibited a reversible MAO-A inhibition displaying up to 60% recovery of enzymatic activity when diluted with substrate (Tyramine). The results of the in silico study indicated docking scores of −9.56 Kcal/mol (C14) and −9.45 Kcal/mol (C6) and exhibited a π-π stacking interaction with the crucial amino acid Trp-397. The compounds were determined to cross the blood–brain barrier (BBB) and displayed favorable gastrointestinal (GI) absorption. Further, the antioxidant assay results demonstrated that the synthesized compounds possess modest free radical scavenging potential. Conclusions: This study displayed the MAO-A inhibitory potential of morpholine-substituted chalcones as a promising pharmacophore for the development of novel antidepressant lead compounds. Full article

Tobacco smoking remains a major global health concern, causing preventable deaths and economic strain. Although new tobacco products such as heat-not-burn (HnB) are safer alternatives to traditional cigarettes, research on their associated risks remains limited. This study aimed to investigate the effects of HnB smoke exposure on the lungs compared to those of traditional cigarettes and the combined use of HnB and cigarettes using experiments with a mouse model. We quantitatively analyzed changes in the levels of 92 blood plasma proteins using the proximity extension assay method and observed significant changes in their levels in mice exposed to different smoke conditions; specifically, the levels of certain proteins, including Ccl20, Cxcl1, and Pdgfb, increased in the HnB smoke-exposed group, suggesting activation of nicotine pathways. Comparative analysis with traditional cigarette smoke-exposed mice further highlighted similarities and differences in their protein expression profiles. This study contributes to an improved understanding of the biological mechanisms underlying the harmful effects of alternative nicotine delivery systems and identifies potential biomarkers associated with the harmful effects of HnB smoke exposure. However, the precise impact of nicotine on the immune system may be influenced by various factors, necessitating further research. Full article

Background: Iron deficiency (ID) often coexists with heart failure (HF), and its prevalence increases with the severity of HF. Intravenous ferric carboxymaltose (FCM) has been associated with improvements in clinical outcomes, functional capacity, and quality of life (QoL) in patients with HF and ID. However, while earlier studies showed favorable results, more recent studies have failed to demonstrate significant improvements in outcomes for patients with heart failure with reduced ejection fraction (HFrEF) and ID. This meta-analysis seeks to provide updated insights into the effectiveness and safety of FCM compared to placebo/standard of care (SoC) among patients with HFrEF and ID/iron deficiency anemia (IDA). Methods: We performed a systematic review and meta-analysis of the literature from inception to December 2023, utilizing databases such as MEDLINE (via PubMed), Google Scholar, the Cochrane Library, ClinicalTrials.gov, and the ScienceDirect portal. A statistical analysis was carried out using RevMan 5.4 with a random-effects model. Dichotomous outcomes were reported as odds ratios (OR), while continuous outcomes were presented as the weighted mean difference (WMD) with corresponding 95% confidence intervals (CI), and heterogeneity was assessed using the I² test. Results: The final analysis included data from six randomized controlled trials (RCTs), comprising 5132 patients. Our findings indicate a significant reduction in total HF hospitalizations among patients with HFrEF and ID/IDA treated with FCM compared to those receiving the placebo or SoC, with an OR of 0.59 (95% CI: 0.40 to 0.88, p < 0.010). However, no statistically significant difference was observed in the total number of deaths between the FCM and placebo/SoC groups (OR: 0.85; 95% CI: 0.70 to 1.03, p = 0.09), non-HF hospitalizations (OR: 0.71; 95% CI: 0.41 to 1.25, p = 0.24), or the composite outcome of cardiovascular hospitalizations and cardiovascular deaths (OR: 0.65; 95% CI: 0.40 to 1.04, p = 0.07). Regarding functional capacity, as assessed by the change in 6-min walk test (6MWT) distance, no significant improvement was found, with a weighted mean difference (WMD) of 14.03 (95% CI: −10.94 to 38.99, p = 0.27). QoL, measured by the Kansas City Cardiomyopathy Questionnaire (KCCQ) score, also did not show significant enhancement, with a WMD of 3.85 (95% CI: −0.55 to 8.24, p = 0.09). Furthermore, the safety analysis revealed no significant difference in the incidence of serious adverse events between the FCM and placebo/SoC groups, with an OR of 0.73 (95% CI: 0.49 to 1.10, p = 0.13). Conclusions: In patients with HFrEF and IDA, treatment with intravenous FCM significantly lowers the risk of total HF hospitalizations but does not appear to affect functional capacity, QoL, or mortality. Full article

The thermal decomposition of a heat-not-burn (HNB) tobacco at four temperatures (250–400 °C) was studied via thermogravimetric analysis (TGA) and Multi-shot pyrolizer experiments (Py-GC/MS), and the effect of four potential additives, USY Beta and beta zeolites and Silica Lovel 6000 and SBA-15 silicates at three concentrations (5, 15 and 25% w/w) under an inert and oxidative atmosphere was analyzed. Different techniques were applied showing that the presence of the additives modifies the decomposition processes (TGA). Py-GC/MS showed that these tobaccos generate large amounts of Nicotine and Glycerine. Acid compounds are the most abundant compounds under an inert atmosphere, while Oxygenated compounds predominate under an oxidative atmosphere. In both atmospheres, Furans and Aromatics present in a significant abundance at high temperatures. The additives used reduce both the number and the concentration of most of the compounds generated, especially at high temperatures and concentrations. Moreover, SBA-15 shows good aptitudes to reduce the formation of some individual compounds included in the FDA’s HPHC list, such as Acetone and Acetaldehyde. Finally, smoking experiments corroborated that all additives produce marked reductions in TPM, i.e., the majority fraction obtained, and in practically all the compounds generated. Phenol, a toxicant compound that was detected in a significant amount, is also markedly reduced. SBA-15 is the material that presents a major reduction in the TPM and the principal compounds generated. These results may be of great interest for further reducing the toxicity of smoking this type of heat-not-burn tobacco product. Full article

Treponema pallidum subspp. pallidum is a spirochaete bacterium that causes syphilis, one of the most common sexually transmitted diseases. Syphilis progresses through four distinct stages, each characterized by specific symptoms, namely primary, secondary, latent, and late (tertiary) syphilis. Serology has been considered the primary diagnostic approach. However, it is plagued by problems such as the limited specificity of nontreponemal tests and the inadequate correlation of treponemal tests with disease activity. In this study, we focused on the development of a loop-mediated isothermal amplification assay utilizing hydroxy naphthol blue (LAMP-HNB) for the diagnosis of T. pallidum subspp. pallidum. Specifically, this study seeks to determine the analytical sensitivity (limit of detection; LOD) and analytical specificity. Four hundred clinical serum samples were analyzed for diagnostic sensitivity, specificity, and predictive value, and each technique’s 95% confidence intervals (95% CI, p < 0.05) were evaluated. The limit of detection for polymerase chain reaction with agarose gel electrophoresis (PCR-AGE), the loop-mediated isothermal amplification assay combined with agarose gel electrophoresis (LAMP-AGE), and LAMP-HNB were 116 pg/µL, 11.6 pg/µL, and 11.6 pg/ µL, respectively. Analytical specificity examinations indicated the absence of cross-reactivity with Leptospira interrogans, Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, human immunodeficiency virus (HIV), and healthy human serum in PCR-AGE, LAMP-AGE, and LAMP-HNB. The diagnostic sensitivity, diagnostic specificity, positive predictive value (PPV), and negative predictive value (NPV) for PCR-AGE were 100.00 (100.00)%, 94.50 (94.40–94.60)%, 94.79 (94.69–94.88)%, and 100.00 (100.00)%, respectively. While, for LAMP-AGE and LAMP-HNB, they were 100.00 (100.00)%, 91.00 (90.87–91.13)%, 91.74 (91.63–91.86)%, and 100.00 (100.00)%, respectively. The LAMP-HNB test is simple, rapid, highly sensitive, and highly specific, without requiring expensive equipment. In the future, the LAMP-HNB assay may develop into a single-step diagnostic process, enabling the use as point-of-care testing for the diagnosis, prevention, and management of syphilis infection. Full article