Urinary thymidine dimer excretion reflects personal ultraviolet radiation exposure levels.

Exposure to ultraviolet radiation (UVR) leads to skin DNA damage, specifically in the form of cyclobutane pyrimidine dimers, with thymidine dimers being the most common. Quantifying these dimers can indicate the extent of DNA damage resulting from UVR exposure. Here, a new liquid chromatography-mass spectrometry (LC-MS) method was used to quantify thymidine dimers in the urine after a temporary increase in real-life UVR exposure. Healthy Danish volunteers (n = 27) experienced increased UVR exposure during a winter vacation. Individual exposure, assessed via personally worn electronic UVR dosimeters, revealed a mean exposure level of 32.9 standard erythema doses (SEDs) during the last week of vacation. Morning urine thymidine dimer concentrations were markedly elevated both 1 and 2 days post-vacation, and individual thymidine dimer levels correlated with UVR exposure during the last week of the vacation. The strongest correlation with erythema-weighted personal UVR exposure (Power model, r2 = 0.64, p < 0.001) was observed when both morning urine samples were combined to measure 48-h thymidine dimer excretion, whereas 24-h excretion based on a single sample provided a weaker correlation (Power model, r2 = 0.55, p < 0.001). Sex, age, and skin phototype had no significant effect on these correlations. For the first time, urinary thymidine dimer excretion was quantified by LC-MS to evaluate the effect of a temporary increase in personal UVR exposure in a real-life setting. The high sensitivity to elevated UVR exposure and correlation between urinary excretion and measured SED suggest that this approach may be used to quantify DNA damage and repair and to evaluate photoprevention strategies.

Development of new matrix reference materials for quantitative urine analysis in drug tests.

Accurate quantitative analyses require standardized methods to control and improve the analytical process in the laboratory. The availability of urine reference materials (RMs) may offer a feasible option to improve the accuracy of urine analysis and to control matrix effects. This paper presents the complete process of the development of matrix RMs in urine, including sample preparation, homogeneity, and stability studies, as well as uncertainty assessment. A freeze-drying process was developed, and freeze-dried human and pig urine samples were prepared and verified to have comparable homogeneity to liquid samples and higher stability than liquid human, pig, and artificial urine samples at 4℃ or room temperature and under extreme conditions. A total of 21 authentic urine samples from August 2022 were measured with freeze-dried RMs and spiked urine samples, and the reliability of the quantification of the RMs was compared. The freeze-dried human urine matrix RM appeared to be an excellent tool for daily quality control, as it showed high stability and gave the most consistent results with spiked samples.

Development of a time-resolved immunochromatographic test strip for rapid and quantitative determination of retinol-binding protein 4 in urine.

Urine retinol-binding protein 4 (RBP4) has recently been reported as a novel earlier biomarker of chronic kidney disease (CKD) which is a global public health problem with high morbidity and mortality. Accurate and rapid detection of urine RBP4 is essential for early monitor of impaired kidney function and prevention of CKD progression. In the present study, we developed a time-resolved fluorescence immunochromatographic test strip (TRFIS) for the quantitative and rapid detection of urine RBP4. This TRFIS possessed excellent linearity ranging from 0.024 to 12.50 ng/mL for the detection of urine RBP4, and displayed a good linearity (Y = 239,581 × X + 617,238, R2 = 0.9902), with the lowest visual detection limit of 0.049 ng/mL. This TRFIS allows for quantitative detection of urine RBP4 within 15 min and shows high specificity. The intra-batch coefficient of variation (CV) and the inter-batch CV were both < 8%, respectively. Additionally, this TRFIS was applied to detect RBP4 in the urine samples from healthy donors and patients with CKD, and the results of TRFIS could efficiently discern the patients with CKD from the healthy donors. The developed TRFIS has the characteristics of high sensitivity, high accuracy, and a wide linear range, and is suitable for rapid and quantitative determination of urine RBP4.

Rapid green analytical methodology for simultaneous monitoring of nitrosamines and semi-volatile organic compounds in water and human urine samples.

Nitrosamines and semi-volatile organic compounds (SVOCs) are carcinogenic contaminants in water and biological matrices. Conventional analytical methods often struggle to detect trace concentrations due to poor extraction efficacies. This study presents a novel, low-cost, in-syringe-assisted fast extraction cum cleanup technique coupled with GC-FID for monitoring four nitrosamines and two SVOCs in drinking water and human urine samples to measure the contamination and exposure levels. This extraction protocol combines a novel green in-syringe liquid-liquid extraction step using dimethyl carbonate as the green extraction solvent, coupled with a semi-automated solid-phase extraction cleanup process. Then, the final extractant is analyzed using gas chromatography-flame ionization detection (GC-FID) for monitoring. The method demonstrated excellent linearity (R2 > 0.998) between 1.5 and 500 ng mL⁻1 for all six target compounds. Detection limits ranged from 1.0 to 2.0 ng mL⁻1. Extraction recoveries were between 87 and 105% for both urine samples and water samples. Intra-day and inter-day precision were below 9% RSD. The blue applicability grade index evaluation scored 70.0, indicating good practical applicability. The developed analytical protocol offers a sensitive, accurate, low-cost, rapid, and environmentally friendly method for simultaneously quantifying multiple nitrosamines and SVOCs in environmental and human samples. Its performance characteristics and sustainability metrics suggest the potential for broad application in monitoring and exposure studies.

Deciphering the human urine matrix: a new approach to simultaneously quantify the main ions and organic compounds by ion chromatography/mass spectrometry (IC-MS).

Analyzing the composition of (human) urine plays a major role in the fields of biology and medicine. Organic molecules (such as urea, creatine) and ions (such as chloride, sulfate) are the major compounds present in urine, the quantification of which allows for the diagnosis of a subject's health condition. Various analytical methods have been reported for studying urine components and validated on the basis of known and referenced compounds. The present work introduces a new method able to simultaneously determine both major organic molecules and ions contained in urine, by combining ion chromatography using a conductimetric detector with mass spectroscopy. The analysis of organic and ionized compounds (anionic and cationic) was achieved in double injections. For quantification, the standard addition method was used. Human urine samples were pre-treated (diluted and filtered) for IC-CD/MS analysis. The analytes were separated in 35 min. Calibration ranges (0-20 mg.L-1) and correlation coefficients (> 99.3%) as well as detection (LODs < 0.75 mg.L-1) and quantification (LOQs < 2.59 mg.L-1) limits were obtained for the main organic molecules (lactic, hippuric, citric, uric, oxalic acids, urea, creatine, and creatinine) and ions (chloride, sulfate, phosphate, sodium, ammonium, potassium, calcium, and magnesium) contained in urine. The intra- and inter-day accuracies of the analytes consistently ranged from 0.1 to 5.0%, and the precision was within 4.0%. For all analytes, no significant matrix effects were observed, and recoveries ranged from 94.9 to 102.6%. Finally, quantitative results of analytes were obtained from 10 different human urine samples.

Fabricating a designer capsule phase microextraction platform based on sol-gel Carbowax 20M-zwitterionic ionic liquid composite sorbent for the extraction of lipid-lowering drugs from human urine samples.

A sol-gel Carbowax 20 M/3-[(3-Cholamidopropyl) dimethyl ammonio]-1-propanesulfonate composite sorbent-based capsule phase microextraction device has been fabricated and characterized for the determination of four statins (pravastatin, rosuvastatin, pitavastatin, and atorvastatin) in human urine. The presence of ionizable carboxyl functional groups in statins requires pH adjustment of the sample matrix to ensure that the target molecules are in their protonated form (pH should be 2 units below their pKa values) which not only is cumbersome but also risks unintended contamination of the sample. This challenge was addressed by introducing zwitterionic ionic liquid in addition to neutral, polar Carbowax 20 M polymer in the sol-gel-derived composite sorbent. As such, the composite zwitterionic multi-modal sorbent can simultaneously extract neutral, cationic, and anionic species. This particular attribute of the composite sorbent eliminates the necessity of the matrix pH adjustment and consequently simplifies the overall sample preparation workflow. Various experimental parameters such as the sample amount, extraction time, salt addition, stirring rate, and elution solvent type that may affect the extraction performance of the statins were investigated using a central composite design and the one-parameter-at-a-time approach. The analytes and the internal standard were separated on a C18 column with gradient elution using phosphate buffer (20 mM, pH 3) and acetonitrile as mobile phase. The analytes were detected at 237 nm. The method was validated, and linearity was observed in the range 0.10-2.0 µg mL-1 for all compounds. The method precision was better 9.9% and 10.4% for intra-day and inter-day, respectively, while the relative recoveries were acceptable, ranging between 83.4 and 116% in all cases. Method greenness was assessed using the ComplexGAPI index. Finally, the method's applicability was demonstrated in the determination of the statins in authentic human urine after oral administration of pitavastatin and rosuvastatin-containing tablets.

Detection of L-cysteine in urine samples based on CdS/TiO2-modified extended-gate field-effect transistor photoelectrochemical sensor.

A novel extended-gate field-effect transistor (FET) photoelectrochemical (EGFET PEC) sensor was designed for highly sensitive detection of L-cysteine (L-Cys). TiO2 was initially modified on the ITO electrode by the sol-gel dip-coating method and calcined to produce TiO2/ITO. Then, CdS was synthesized on the TiO2 surface by hydrothermal method to obtain the CdS-TiO2 heterojunction material. CdS/TiO2/ITO was connected to the gate of the FET to obtain an EGFET PEC sensor. Under the irradiation of a xenon lamp simulating visible light, the CdS/TiO2 heterojunction composite absorbs light energy to produce photogenerated electron-hole pairs, which have strong photocatalytic oxidation activity and oxidize L-Cys covalently identified by Cd(II) through CdS covalent. These pairs generate a photovoltage that controls the current between the source and the drain to detect L-Cys. Under the optimized experimental conditions, the optical drain current (ID) of the sensor exhibited a good linear relationship with the logarithm of L-Cys in the range of 5.0 × 10-9-1.0 × 10-6 mol/L, and the detection limit was 1.3 × 10-9 mol/L (S/N = 3), which is lower than the values reported by other detection methods. Results showed that the CdS/TiO2/ITO EGFET PEC sensor revealed high sensitivity and good selectivity. The sensor has been used to determine L-Cys in urine samples.

Low-Cost Electronic Nose for the Determination of Urinary Infections.

Currently, urine samples for bacterial or fungal infections require a long diagnostic period (48 h). In the present work, a point-of-care device known as an electronic nose (eNose) has been designed based on the "smell print" of infections, since each one emits various volatile organic compounds (VOC) that can be registered by the electronic systems of the device and recognized in a very short time. Urine samples were analyzed in parallel using urine culture and eNose technology. A total of 203 urine samples were analyzed, of which 106 were infected and 97 were not infected. A principal component analysis (PCA) was performed using these data. The algorithm was initially capable of correctly classifying 49% of the total samples. By using SVM-based models, it is possible to improve the accuracy of the classification up to 74% when randomly using 85% of the data for training and 15% for validation. The model is evaluated as having a correct classification rate of 74%. In conclusion, the diagnostic accuracy of the eNose in urine samples is high, promising and amenable for further improvement, and the eNose has the potential to become a feasible, reproducible, low-cost and high-precision device to be applied in clinical practice for the diagnosis of urinary tract infections.

Sensing of Catecholamine in Human Urine Using a Simple Colorimetric Assay Based on Direct Melanochrome and Indolequinone Formation.

We used the first enzyme-free synthesis and stabilization of soluble melanochrome (MC) and 5,6-indolequinone (IQ) derived from levodopa (LD), dopamine (DA), and norepinephrine (NE) oxidation to develop a simple colorimetric assay for catecholamine detection in human urine, also elucidating the time-dependent formation and molecular weight of MC and IQ using UV-Vis spectroscopy and mass spectrometry. The quantitative detection of LD and DA was achieved in human urine using MC as a selective colorimetric reporter to demonstrate the potential assay applicability in a matrix of interest in therapeutic drug monitoring (TDM) and in clinical chemistry. The assay showed a linear dynamic range between 5.0 mg L-1 and 50.0 mg L-1, covering the concentration range of DA and LD found in urine samples from, e.g., Parkinson's patients undergoing LD-based pharmacological therapy. The data reproducibility in the real matrix was very good within this concentration range (RSDav% 3.7% and 6.1% for DA and LD, respectively), also showing very good analytical performances with the limits of detection of 3.69 ± 0.17 mg L-1 and 2.51 ± 0.08 mg L-1 for DA and LD, respectively, thus paving the way for the effective and non-invasive monitoring of dopamine and levodopa in urine from patients during TDM in Parkinson's disease.

Serotonin-Derived Fluorophore: A Novel Fluorescent Biomaterial for Copper Detection in Urine.

We took advantage of the fluorescent features of a serotonin-derived fluorophore to develop a simple and low-cost assay for copper in urine. The quenching-based fluorescence assay linearly responds within the concentration range of clinical interest in buffer and in artificial urine, showing very good reproducibility (CVav% = 4% and 3%) and low detection limits (16 ± 1 µg L-1 and 23 ± 1 µg L-1). The Cu2+ content was also estimated in human urine samples, showing excellent analytical performances (CVav% = 1%), with a limit of detection of 59 ± 3 µg L-1 and a limit of quantification of 97 ± 11 µg L-1, which are below the reference value for a pathological Cu2+ concentration. The assay was successfully validated through mass spectrometry measurements. To the best of our knowledge, this is the first example of copper ion detection exploiting the fluorescence quenching of a biopolymer, offering a potential diagnostic tool for copper-dependent diseases.

Automated Urine Cell Image Classification Model Using Chaotic Mixer Deep Feature Extraction.

Microscopic examination of urinary sediments is a common laboratory procedure. Automated image-based classification of urinary sediments can reduce analysis time and costs. Inspired by cryptographic mixing protocols and computer vision, we developed an image classification model that combines a novel Arnold Cat Map (ACM)- and fixed-size patch-based mixer algorithm with transfer learning for deep feature extraction. Our study dataset comprised 6,687 urinary sediment images belonging to seven classes: Cast, Crystal, Epithelia, Epithelial nuclei, Erythrocyte, Leukocyte, and Mycete. The developed model consists of four layers: (1) an ACM-based mixer to generate mixed images from resized 224 × 224 input images using fixed-size 16 × 16 patches; (2) DenseNet201 pre-trained on ImageNet1K to extract 1,920 features from each raw input image, and its six corresponding mixed images were concatenated to form a final feature vector of length 13,440; (3) iterative neighborhood component analysis to select the most discriminative feature vector of optimal length 342, determined using a k-nearest neighbor (kNN)-based loss function calculator; and (4) shallow kNN-based classification with ten-fold cross-validation. Our model achieved 98.52% overall accuracy for seven-class classification, outperforming published models for urinary cell and sediment analysis. We demonstrated the feasibility and accuracy of deep feature engineering using an ACM-based mixer algorithm for image preprocessing combined with pre-trained DenseNet201 for feature extraction. The classification model was both demonstrably accurate and computationally lightweight, making it ready for implementation in real-world image-based urine sediment analysis applications.

Target amplification-free detection of urinary microRNA for diabetic nephropathy diagnosis with electrocatalytic reaction.

Diabetic nephropathy (DN) is a serious diabetic complication, usually developed from type II diabetes mellitus (T2DM) and known as type II DN (T2DN). New emerging biomarkers for T2DN are microRNAs (miRNAs) which have been studied for the noninvasive early-stage detection of the disease. In this work, a nucleic acid amplification-free miRNA-124 sensor based on target-induced strand displacement on magnetic beads, and by using methylene blue-loaded silica particles as a label was developed. Measurement methods can be either visual observation, spectrophotometry, or electrochemistry. After incubation and separation of the magnetic particles, a blue-violet solution (564 nm) appeared, depending on the concentration of miRNA displaced. For electrochemical detection, methylene blue on the silica served as a redox mediator for the coupled reaction with ferricyanide in the solution phase. At the electrode surface, ferricyanide was re-reduced to ferrocyanide, and was thus available for further reaction with methylene blue, forming an amplification cycle. After optimization, the total assay time was 60 min, and limits of detection were 1 pM, 6 fM, and 0.65 fM, by the naked eye, spectrophotometry and electrochemistry, respectively. The miRNAs in 42 suspected urine samples from patients suffering from either diabetic nephropathy, diabetes mellitus, or chronic kidney disease were validated by comparing with the droplet digital polymerase chain reaction (ddPCR).

Comprehensive suspect screening for the identification of contaminants of emerging concern in urine of Flemish adolescents by liquid chromatography high-resolution mass spectrometry.

The increasing human exposure to contaminants of emerging concern (CECs) cannot be fully assessed by targeted biomonitoring methods alone as these are limited to a subset of known analytes. On the contrary, suspect screening approaches based on liquid chromatography coupled to high-resolution mass spectrometry (LC-HRMS) allow the simultaneous detection of a high number of CECs and/or their (predicted) metabolites leading to a more comprehensive assessment of possible human exposure to these compounds. Within this study, 83 urine samples of Flemish adolescents (47 males, 36 females) collected in the frame of the 4th cycle of the Flemish Environment and Health Study (FLEHS IV) were selected with the aim of including a high and a low exposure group based on the overall exposure of 45 known contaminants. Samples were analyzed using a previously developed method involving a suspect screening approach to annotate CECs and their metabolites. The applied suspect list contained a total of >12,500 CECs and their known and predicted metabolites resulting from metabolization reactions, such as hydroxylation, glucuronidation and methylation. In total, 63 compounds were annotated at a confidence level of 3 or better, with most of the detected compounds not included in current biomonitoring programs. 5 out of the 63 compounds could be assigned with confidence level 2. Five compounds could unequivocally be identified (confidence level 1) through the comparison with reference standards. Personal care products were the main detected compound class (42% of detected compounds). Additionally, a detailed literature search indicated potential toxic effects for several of the detected CECs. Lastly, in the urine samples, a significantly higher number (p < 0.05) of compounds was detected in the high exposure group as opposed to the low exposure group. This difference could only be observed between high and low exposure load samples of female participants (p < 0.01).

Current applications of capillary electrophoresis-mass spectrometry for the analysis of biologically important analytes in urine (2017 to mid-2021): A review.

Capillary electrophoresis coupled online with mass detection is a modern tool for analyzing wide ranges of compounds in complex samples, including urine. Capillary electrophoresis with mass spectrometry allows the separation and identification of various analytes spanning from small ions to high molecular weight protein complexes. Similarly to the much more common liquid chromatography-mass spectrometry techniques, the capillary electrophoresis separation reduces the complexity of the mixture of analytes entering the mass spectrometer resulting in reduced ion suppression and a more straightforward interpretation of the mass spectrometry data. This review summarizes capillary electrophoresis with mass spectrometry studies published between the years 2017 and 2021, aiming at the determination of various compounds excreted in urine. The properties of the urine, including its diagnostical and analytical features and chemical composition, are also discussed including general protocols for the urine sample preparation. The mechanism of the electrophoretic separation and the instrumentation for capillary electrophoresis with mass spectrometry coupling is also included. This review shows the potential of the capillary electrophoresis with mass spectrometry technique for the analyses of different kinds of analytes in a complex biological matrix. The discussed applications are divided into two main groups (capillary electrophoresis with mass spectrometry for the determination of drugs and drugs of abuse in urine and capillary electrophoresis with mass spectrometry for the studies of urinary metabolome).

Peptidomic Analysis of Urine from Youths with Early Type 1 Diabetes Reveals Novel Bioactivity of Uromodulin Peptides In Vitro.

Chronic hyperglycemia is known to disrupt the proteolytic milieu, initiating compensatory and maladaptive pathways in the diabetic kidney. Such changes in intrarenal proteolysis are captured by the urinary peptidome. To elucidate the early kidney response to chronic hyperglycemia, we conducted a peptidomic investigation into urines from otherwise healthy youths with type 1 diabetes and their non-diabetic peers using unbiased and targeted mass spectrometry-based techniques. This cross-sectional study included two separate cohorts for the discovery (n = 30) and internal validation (n = 30) of differential peptide excretion. Peptide bioactivity was predicted using PeptideRanker and subsequently verified in vitro Proteasix and the Nephroseq database were used to identify putative proteases responsible for peptide generation and examine their expression in diabetic nephropathy. A total of 6550 urinary peptides were identified in the discovery analysis. We further examined the subset of 162 peptides, which were quantified across all thirty samples. Of the 15 differentially excreted peptides (p < 0.05), seven derived from a C-terminal region (589SGSVIDQSRVLNLGPITRK607) of uromodulin, a kidney-specific protein. Increased excretion of five uromodulin peptides was replicated in the validation cohort using parallel reaction monitoring (p < 0.05). One of the validated peptides (SGSVIDQSRVLNLGPI) activated NFκB and AP-1 signaling, stimulated cytokine release, and enhanced neutrophil migration in vitro. In silico analyses highlighted several potential proteases such as hepsin, meprin A, and cathepsin B to be responsible for generating these peptides. In summary, we identified a urinary signature of uromodulin peptides associated with early type 1 diabetes before clinical manifestations of kidney disease and discovered novel bioactivity of uromodulin peptides in vitro Our present findings lay the groundwork for future studies to validate peptide excretion in larger and broader populations, to investigate the role of bioactive uromodulin peptides in high glucose conditions, and to examine proteases that cleave uromodulin.

An in-depth Comparison of the Pediatric and Adult Urinary N-glycomes.

We performed an in-depth characterization and comparison of the pediatric and adult urinary glycomes using a nanoLC-MS/MS based glycomics method, which included normal healthy pediatric (1-10 years, n = 21) and adult (21-50 years, n = 22) individuals. A total of 116 N-glycan compositions were identified, and 46 of them could be reproducibly quantified. We performed quantitative comparisons of the 46 glycan compositions between different age and sex groups. The results showed significant quantitative changes between the pediatric and adult cohorts. The pediatric urinary N-glycome was found to contain a higher level of high-mannose (HM), asialylated/afucosylated glycans (excluding HM), neutral fucosylated and agalactosylated glycans, and a lower level of trisialylated glycans compared with the adult. We further analyzed gender-associated glycan changes in the pediatric and adult group, respectively. In the pediatric group, there was almost no difference of glycan levels between males and females. In adult, the majority of glycans were more abundant in males than females, except the high-mannose and tetrasialylated glycans. These findings highlight the importance to consider age-matching and adult sex-matching for urinary glycan studies. The identified normal pediatric and adult urinary glycomes can serve as a baseline reference for comparisons to other disease states affected by glycosylation.

Characterizing Patients with Recurrent Urinary Tract Infections in Vesicoureteral Reflux: A Pilot Study of the Urinary Proteome.

Recurrent urinary tract infections (UTIs) pose a significant burden on the health care system. Underlying mechanisms predisposing children to UTIs and associated changes in the urinary proteome are not well understood. We aimed to investigate the urinary proteome of a subset of children who have vesicoureteral reflux (VUR) and recurrent UTIs because of their risk of developing infection-related renal damage. Improving diagnostic modalities to identify UTI risk factors would significantly alter the clinical management of children with VUR. We profiled the urinary proteomes of 22 VUR patients with low grade VUR (1-3 out of 5), a history of recurrent UTIs, and renal scarring, comparing them to those obtained from 22 age-matched controls. Urinary proteins were analyzed by mass spectrometry followed by protein quantitation based on spectral counting. Of the 2,551 proteins identified across both cohorts, 964 were robustly quantified, as defined by meeting criteria with spectral count (SC) ≥2 in at least 7 patients in either VUR or control cohort. Eighty proteins had differential expression between the two cohorts, with 44 proteins significantly up-regulated and 36 downregulated (q <0.075, FC ≥1.2). Urinary proteins involved in inflammation, acute phase response (APR), modulation of extracellular matrix (ECM), and carbohydrate metabolism were altered among the study cohort.

Diagnostic performance of urine analysis based on flow microimaging and artificial intelligence recognition technology in suspected urinary tract infection patients.

To investigate the diagnostic performance and clinical value of flow microimaging and artificial intelligence recognition technology for rapid screening of suspected urinary tract infection (UTI). Standard strains of bacteria responsible for UTIs, Escherichia coli ATCC25922 and Staphylococcus aureus ATCC25923 were prepared and used to evaluate the accuracy of classifying and counting bacteria. A total of 146 specimens of clean, midstream urine were collected from adults with suspected UTI (excluding pregnant women, and patients with urethral catheterization) and analyzed by urinary culture and assay using a MUS-3600 analyzer. Fourfold tables were used to evaluate the diagnostic performance in measurements of nitrite and leukocyte esterase. ROC curves were generated to identify cutoff values for bacillus, coccus, leukocyte, and leukocyte cluster counts. Of the samples cultured, 85 (58%) were negative and 61 (42%) were positive. E. coli and S. aureus showed good linear relationships between measured and theoretical values in a series of standard strains samples (R2>0.95). The sensitivity of the nitrite parameter for diagnosis of suspected UTI was 37.7% and the specificity was 100%. Cutoff values obtained from ROC analysis were 50 µl for bacillus (sensitivity: 69.5%; specificity: 96.5%) with positive predictive value of 93.2% and negative predictive value of 82%. WBC ≥ 23 µl or bacillus ≥50 µl gave the highest sensitivity of 81% and NPV of 86%. MUS-3600 demonstrated sufficient specificity for UTIs to have utility in reducing needless urinary culture. Accuracy in classifying bacillus shows great potential for the rapid identification of pathogens for clinical purposes.

Effects of a new 1,2,3-thiadiazole containing hydrazone antimycobacterial agent on serum and liver biochemical parameters in female mice.

Isoniazid (INH), a first-line drug in anti-tuberculosis therapy, is known to be potentially harmful and is associated with numerous side effects especially in the blood and liver. In the course of our previous investigations, 1,2,3-thiadiazole containing hydrazone (compound 3) showed excellent antimycobacterial activity against a referent strain M. tuberculosis H37Rv (MIC value 0.39 µM), low cytotoxicity, and did not have toxic effects when administered by oral or intraperitoneal routes to experimental animals (selectivity index SI > 1979, LD50>2000 mg/kg b.w.) what revealed its suitability for further exploration. In the present study compound 3 was chosen to determine its effects on the liver and kidney functions in female mice. The compound was administered orally for 14 days at three doses (100, 200, and 400 mg/kg b.w.). The quantity of malondialdehyde (MDA), the level of reduced glutathione (GSH), blood hematological and biochemical parameters were assessed, and urine analysis was carried out. As a positive control INH was used orally at a dose of 50 mg/kg b.w. The investigated compound 3 did not affect the urine and serum hematological and biochemical parameters as INH did, compared to those of the control mice. The new compound did not affect significantly the MDA quantity and maintained its level near to the control values, though lower by 36% (p < 0.05) than in the INH treated animals. At the higher doses, 200 and 400 mg/kg, it depleted the GSH content by 25% (p < 0.05), compared to the control. However, its level remained 47% (p < 0.05) higher than in the INH treated animals.

Carbonized π-conjugated polymer-coated porous silica: preparation and evaluating its extraction ability for berberine.

In view of the limitations of existing berberine solid-phase extraction adsorbents, this paper proposes a novel carbonized π-conjugated polymer-coated porous silica (SiO2@C-π-CP) adsorbent with simple process and low cost for efficient extraction of berberine by multiple interactions. Characterization methods, including Brunner-Emmet-Teller measurement, thermogravimetric analysis, X-ray photoelectron spectroscopy, and scanning electron microscopy techniques, were used to verify the successful modification of carbonized π-conjugated polymer on the surface of porous silica. The berberine was selected as target molecule, and the adsorption mechanism and process were investigated through adsorption kinetics, adsorption isotherms, and thermodynamic studies. The fitting results show that the adsorption of berberine by SiO2@C-π-CP well conforms to the pseudo-second-order and Langmuir models. By optimizing the main SPE parameters, the SPE method based on SiO2@C-π-CP was developed. Excellent results were obtained, including low limit of detection (0.75 ng mL-1) and limit of quantification (2 ng mL-1), wide linearity (2-13,000 ng mL-1), and satisfactory relative standard deviations (RSD) of inter-day (1.5%) and intra-day (6.2%). Finally, the SiO2@C-π-CP also has been successfully used to the enrichment of berberine in real urine samples. This research makes clear that SiO2@C-π-CP has outstanding potential for trace enrichment of berberine alkaloids.