Body Fluids Research Articles

RNA Analysis in Forensic Molecular Biology.

Different types of RNA take on multiple crucial functions in living cells and tissues. Messenger RNA (mRNA) is a temporary molecular carrier of genetic information. Analysis of the composition of all mRNA contained in a cell at a given moment, the so-called transcriptome, enables the determination of the type of cell and its condition, e.g., in pathologically altered states. This review is based on pertinent publications retrieved by a selective literature search. The analysis of differential gene expression has already been used in forensic molecular biology to determine the type of tissue contained in biological specimens. It is also being used in criminal investigations to determine the composition of mixed traces of various bodily fluids and/or organ tissues. The method is limited by degradation of the mRNA molecules through environmental influences. The use of newly developed molecular biological methods such as massive parallel sequencing can expand the information obtainable by this investigative method. Current research also addresses the forensic potential of deriving relevant information about the crime-e.g., its timing, or the condition of the involved persons-from the totality of mRNA species present in the specimens. Forensic RNA analysis can yield a great deal of relevant information. It is likely to be applicable in a much wider variety of forensic situations in the near future.

Reference genes to normalize body fluid identification data obtained using RT-qPCR

ABSTRACT The use of messenger RNA for body fluid identification has been well documented. While reference genes are commonly used for normalizing gene expression, the use of reference genes in forensic science is not straightforward. Degradation of samples, mixed body fluids, and small volumes contribute to the complexity of normalizing gene expression. To demonstrate normalization of gene expression for forensic body fluid identification, a preliminary study involving two reference genes (beta-2-microglobulin, B2M and glucose-6-phosphate dehydrogenase, G6PD) was undertaken. Novel reverse-transcription quantitative PCR (RT-qPCR) assays for circulatory blood, buccal swabs, menstrual fluid, spermatozoa, seminal fluid, and vaginal material were designed. An appropriate PCR efficiency for the B2M and G6PD reference gene assays across the body fluid panel was confirmed prior to assessing their expression levels. B2M was found to be more highly expressed across the different body fluids than G6PD. The normalization of target assay expression using both genes was investigated. However, high variation and poor sensitivity, particularly in buccal and semen samples, were observed for both, which would limit sensitivity and mixture detection in forensic casework. If CT values are the intended output rather than comparative quantification, we propose that reference gene normalization is not required and could be detrimental to interpretation.

Analytical applications of droplet deposition Raman spectroscopy.

The droplet deposition methods in Raman spectroscopy have received considerable attention in the field of analytical sensing focusing on effective pre-concentration of the studied analyte (coffee-ring effect or small spots). This review covers different analytical applications of drop-coating deposition Raman scattering (DCDRS) and droplet deposition surface-enhanced Raman scattering (SERS) spectroscopy. Two main advantages of droplet deposition Raman techniques are considered: the drying-induced segregation of the components from the mixtures (such as body fluids) and the sensitivity of detection of various analytically important molecules. Some recent advanced applications, including clinical cancer diagnosis, are discussed and summarized. Finally, the potential and further perspectives of the droplet deposition Raman methods for analytical studies are introduced.

Serum neurofilament light chain as a prognostic marker of all-cause mortality in a national sample of US adults.

Neurofilament light chain (NfL) is a neuron-specific structural protein released into the extracellular space, including body fluids, upon neuroaxonal damage. Despite evidence of a link in neurological disorders, few studies have examined the association of serum NfL with mortality in population-based studies. Data from the National Health and Nutrition Survey were utilized including 2,071 Non-Hispanic White, Non-Hispanic Black and Hispanic adult participantsand adult participants of other ethnic groups (20-85 years) with serum NfL measurements who were followed for ≤ 6 years till 2019. We tested the association of serum NfL with mortality in the overall population and stratified by sex with the addition of potential interactive and mediating effects of cardio-metabolic risk factors and nutritional biomarkers. Elevated serum NfL levels (above median group) were associated with mortality risk compared to the below median NfL group in the overall sample (P = 0.010), with trends observed within each sex group (P < 0.10). When examining Loge NfL as a continuum, one standard deviation of Loge NfL was associated with an increased mortality risk (HR = 1.88, 95% CI 1.60-2.20, P < 0.001) in the reduced model adjusted for age, sex, race, and poverty income ratio; a finding only slightly attenuated with the adjustment of lifestyle and health-related factors. Four-way decomposition indicated that there was, among others, mediated interaction between NfL and HbA1c and a pure inconsistent mediation with 25(OH)D3 in predicting all-cause mortality, in models adjusted for all other covariates. Furthermore, urinary albumin-to-creatinine ratio interacted synergistically with NfL in relation to mortality risk both on the additive and multiplicative scales. These data indicate that elevated serum NfL levels were associated with all-cause mortality in a nationally representative sample of US adults.

Microplastics: Omnipresent and an ongoing challenge for medical science.

Micro- and nanoplastics are omnipresent not only in the environment, but have also been detected in human body fluids and tissue. The subsequent commentary provides a perspective about potential risks for human health as well as resulting challenges for medical science.

Assessing the reduction of viral infectivity in HPV16/18-positive women after one, two, and three doses of Gardasil-9 (RIFT): Study protocol.

Human Papillomavirus (HPV) prophylactic vaccination has proven effective in preventing new infections, but it does not treat existing HPV infections or associated diseases. Hence, there is still an important reservoir of HPV in adults, as vaccination programs are mainly focused on young women. The primary objective of this non-randomized, open-label trial is to evaluate if a 3-dose regimen of Gardasil-9 in HPV16/18-positive women could reduce the infective capacity of their body fluids. We aim to assess if vaccine-induced antibodies could neutralize virions present in the mucosa, thus preventing the release of infective particles and HPV transmission to sexual partners. As our main endpoint, the E1^E4-HaCaT model will be used to assess the infectivity rate of cervical, anal and oral samples, obtained from women before and after vaccination. HPV DNA positivity, virion production, seroconversion, and the presence of antibodies in the exudates, will be evaluated to attribute infectivity reduction to vaccination. Our study will recruit two different cohorts (RIFT-HPV1 and RIFT-HPV2) of non-vaccinated adult women. RIFT-HPV1 will include subjects with an HPV16/18 positive cervical test and no apparent cervical lesions or cervical lesions eligible for conservative treatment. RIFT-HPV2 will include subjects with an HPV16/18 positive anal test and no apparent anal lesions or anal lesions eligible for conservative treatment, as well as women with an HPV16/18 positive cervical test and HPV-associated vulvar lesions. Subjects complying with inclusion criteria for both cohorts will be recruited to the main cohort, RIFT-HPV1. Three doses of Gardasil-9 will be administered intramuscularly at visit 1 (0 months), visit 2 (2 months) and visit 3 (6 months). Even though prophylactic HPV vaccines would not eliminate a pre-existing infection, our results will determine if HPV vaccination could be considered as a new complementary strategy to prevent HPV-associated diseases by reducing viral spread. Trial registration: https://clinicaltrials.gov/ct2/show/NCT05334706.

Relevant MicroRNAs of MMPs and TIMPs with Certain Gut Microbiota Could Be Involved in the Invasiveness and Metastasis of Malignant Tumors

Malignant tumors are heterogeneous diseases characterized by uncontrolled cell proliferation, invasion, and/or metastasis. In particular, cancer stem cells within these tumors might be responsible for the invasiveness and the property of recurrence of malignancies. It has been reported that matrix metalloproteases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) could contribute to the development of tumor invasiveness. Remarkably, specific MMPs can drive the malignant phenotype of tumors counting the acquisition of stem cell feature, in which STATs signaling might play an important role. Furthermore, MMPs might be crucial for extra-cellular matrix remodeling in the pathological condition of cancer stem cells. Some immune responses against cancer stem cells in the connection to MMPs and/or TIMPs have been revealed, which might be an important issue in the treatment of aggressive cancers. MicroRNAs (miRNAs) have emerged as favorable biomarkers owing to their important roles in gene regulation and/or stability. Differential expression patterns of specific miRNA may be associated with different types of tumor. In addition, miRNAs can be detected in various body fluids, proposing easily available specimens for diagnostic purposes in tumor microenvironment. Remarkably, it has been shown that miRNA expression might be also related with the gut microbiota composition. Early detection and accurate monitoring of tumors are critical for effective treatment and/or improved patient outcomes. A better understanding of this mechanism of epigenetics could be of use to bring about innovative tactics for the treatment of malignant tumors.

Diagnostic and prognostic significance of mast cell markers in HIV/AIDS: Current insights and future directions.

Human immunodeficiency virus (HIV) infection continues to pose significant global health challenges, necessitating advancements in diagnostic and prognostic approaches to optimize disease management. While primarily recognized for their roles in allergic responses, mast cells have emerged as potential markers with diagnostic and prognostic significance in the context of HIV/AIDS. This paper aims to synthesize current insights and delineate future directions regarding the utility of mast cell markers in diagnosing HIV infection, predicting disease progression, and guiding therapeutic strategies. Mast cells, equipped with distinct markers such as tryptase, chymase, carboxypeptidase A3, and c-kit/CD117 receptors, exhibit tissue-specific expression patterns that offer potential as diagnostic indicators for HIV infection. Understanding the dynamics of these markers in different tissues and body fluids holds promise for accurate HIV diagnosis, disease staging, and monitoring treatment responses. Moreover, the prognostic significance of mast cell markers in HIV/AIDS lies in their potential to predict disease progression, immune dysregulation, and clinical outcomes. The integration of mast cell markers into clinical applications offers promising avenues for refining diagnostic assays, patient monitoring protocols, and therapeutic strategies in HIV/AIDS. Future research directions involve the development of novel diagnostic tools and targeted therapies based on mast cell-specific markers, potentially revolutionizing clinical practice and enhancing patient care in the management of HIV/AIDS. Continued investigations into mast cell markers' diagnostic and prognostic implications hold immense potential to advance our understanding and improve outcomes in HIV/AIDS management.

A Review on High Performance Liquid Chromatography (HPLC)

High-performance liquid chromatography (HPLC) is an important qualitative and quantitative technique, generally used for the estimation of pharmaceutical and biological samples. It is the most versatile, safest, dependable, and fastest chromatographic technique for the quality control of drug components. This article was prepared with a review different aspects of HPLC, such as principle types, instrumentation, and application. High- Performance Liquid Chromatography has played a significant role in clinical laboratories for the separation and quantitation of biomarkers in different body fluids. The development of HPLC involves four basic steps; scouting, optimization, robustness testing, and validation. The technique is used to analyze drugs and medicines for their purity and to maintain the highest standards for pharmaceutical products with the end goal of helping patients with medical issues. Method of validation is the process used to confirm that the analytical procedure employed for a specific test is suitable for its intended use. Validation High-Performance Liquid Chromatography method as per ICH guidelines covers all the performance characteristics of validation, like Accuracy, precision, specificity, linearity, range, limit of detection, limit of quantification, robustness, system suitability the testing. The limitation of High-Performance Liquid Chromatography methods of development, public health importance, and validation is the automated process becomes complicated, has low separation power, and is expensive but High-Performance Liquid Chromatography is the modern diagnostic technique is used in all sectors

Advanced Biomimetic and Biohybrid Magnetic Micro/Nano‐Machines

AbstractBiomimetic and biohybrid micro/nano‐structures involve the replication and creation of technologies, structures, and materials based on biological systems at the micrometer and nanometer scale. These strategies harness the natural biological principles to develop innovative treatment methods and advanced microstructure devices for noninvasive therapies. In this study, a detailed overview of fabrication processes, magnetically assisted locomotive techniques, and potential applications of biomimetic and biohybrid micro/nano‐machines are presented. The latest advancements in magnetically actuated biomimetic structures, such as annelid‐worm‐like microswimmers, jellyfish‐shaped microparticles, fish‐shaped microswimmers, and walnut‐shaped micromotors are explored. Additionally, the magnetic biohybrid systems, including sunflower seed‐based micro‐perforators, nanomotors extracted from the bamboo stem, sperm cell‐based micromotors, bacteria‐based robots, scaffold‐based microrobots, DNA‐based micromotors, microalgae‐based microswimmers, and red blood cell‐based microswimmers are also examined. A thorough investigation of the magnetically assisted locomotive behavior of these microstructure devices in biological Newtonian fluids, featuring cork‐screw motion, undulatory motion, surface wrinkling motion, traveling wave‐like motion, and ciliary stroke motion is discussed. Furthermore, unprecedented and innovative treatment methods developed using these minuscule devices such as cervical cancer treatment using tetrapod hybrid sperm micromotors, tissue regeneration using silk fibroin protein‐based magnetic microscale scaffolds, and doxorubicin drug delivery using mushroom‐based microrobots is extensively presented.

Multi-targeted nanoarrays for early broad-spectrum detection of lung cancer based on blood biopsy of tumor exosomes

Liquid biopsies utilizing tumor exosomes offer a noninvasive approach for cancer diagnosis. However, validation studies consistently report that in the early stages of cancer, the secretion of exosomes by cancer cells is relatively low, while bodily fluids exhibit a high abundance of other interfering biomolecules. Additionally, target mutations or differences in biomarker expression among various lung cancer subtypes may contribute to detection failures. In this study, we propose a targeted nanoarray-based early cancer diagnostic approach for multiple subtypes of lung cancer. The targeted nanoarray was constructed by modifying five targeting aptamers onto mesoporous silica nanoparticles through the conjugation between amino and carboxyl groups. The flow cytometry experiments demonstrated the specific recognition ability of the targeted nanoarray to tumor exosomes in PBS, even at biomarker expression levels as low as 1.5%. Moreover, the TEM results indicated that the targeted nanoarray could isolate tumor exosomes in the blood of tumor-bearing mice. Furthermore, the targeted nanoarray could detect tumor exosomes in the blood of various lung cancer bearing mice, including at the early stages of cancer, which has just been established for 7 days. Overall, the targeted nanoarray represents a promising tool for the early detection of various subtypes of lung cancer.

Loading of antibiotic molecules into organically modified layered calcium silicate

Loading of antibiotic molecules into organically modified layered calcium silicate

Sero-prevalence of hepatitis B virus and associated factors among pregnant women attending antenatal care in Bosaso general hospital, Puntland, Somalia 2023: A cross-sectional study design

Background: Hepatitis is an inflammation of the liver caused by a variety of infectious and non-infectious agents, leading to a range of health problems. Hepatitis B virus (HBV) can cause both acute and chronic liver disease, and the most common virus that can be transmitted from mother to child and other modes of transmission are contact with blood or other body fluids during sexual intercourse, unsafe injections, or exposure to sharp instruments. Objective: To determine the seroprevalence and associated factors of HBV infection among pregnant women attending the Antenatal Clinic at Bosaso General Hospital, Puntland, Somalia, in 2023. Methods: A cross-sectional study was conducted among 302 pregnant women who underwent routine ANC at Bosaso General hospital from March 15 to May 15. Study participants were interviewed using a structured questionnaire. A systematic random sampling technique was used to identify participants. Data entry, editing, and cleanup of the collected data were performed by the principal investigator using EPI data, and data analysis was performed using SPSS software version 20 to examine the association between each independent variable and the outcome variable. Bivariate and multivariate logistic regression analyses were performed to determine associations. Result: A total of 302 respondents participated in the study, with a response rate of 96.8%. The overall prevalence of hepatitis B infection among pregnant women was 6.6% (95% CI = 4.0–9.8). Having a history of dental procedures (AOR = 5.52, 95% CI = 1.190–25.68), a history of blood transfusion (AOR = 5.02, 95% CI = 1.088–23.19), a history surgical procedure (AOR = 4.7, 95% CI = 1.014–21.76), or having a family history of HBV infection (AOR = 10.07, 95% CI = (1.27–79.63) were significant predictors of HBV infection among pregnant women. Conclusion: The seroprevalence of HBV was intermediate (2–8%) in the study area according to WHO classification. A history of dental procedures, blood transfusion, surgical procedures, and a family history of hepatitis B virus infection were significantly associated with HBV infection. Keywords: Sero-prevalence; Bosaso; hepatitis B virus; pregnant women.

Highly Stable Solid Contact Calcium Ion-Selective Electrodes: Rapid Ion-Electron Transduction Triggered by Lipophilic Anions Participating in Redox Reactions of CunS Nanoflowers.

Solid contact (SC) calcium ion-selective electrodes (Ca2+-ISEs) have been widely applied in the analysis of water quality and body fluids by virtue of the unique advantages of easy operation and rapid response. However, the potential drift during the long-term stability test hinders their further practical applications. Designing novel redox SC layers with large capacitance and high hydrophobicity is a promising approach to stabilize the potential stability, meanwhile, exploring the transduction mechanism is also of great guiding significance for the precise design of SC layer materials. Herein, flower-like copper sulfide (CunS-50) composed of nanosheets is meticulously designed as the redox SC layer by modification with the surfactant (CTAB). The CunS-50-based Ca2+-ISE (CunS-50/Ca2+-ISE) demonstrates a near-Nernstian slope of 28.23 mV/dec for Ca2+ in a wide activity linear range of 10-7 to 10-1 M, with a low detection limit of 3.16 × 10-8 M. CunS-50/Ca2+-ISE possesses an extremely low potential drift of only 1.23 ± 0.13 μV/h in the long-term potential stability test. Notably, X-ray absorption fine-structure (XAFS) spectra and electrochemical experiments are adopted to elucidate the transduction mechanism that the lipophilic anion (TFPB-) participates in the redox reaction of CunS-50 at the solid-solid interface of ion-selective membrane (ISM) and redox inorganic SC layer (CunS-50), thereby promoting the generation of free electrons to accelerate ion-electron transduction. This work provides an in-depth comprehension of the transduction mechanism of the potentiometric response and an effective strategy for designing redox materials of ion-electron transduction triggered by lipophilic anions.

AAV-mediated expression of mouse or human GLDC normalises metabolic biomarkers in a GLDC-deficient mouse model of Non-Ketotic Hyperglycinemia

Non-Ketotic Hyperglycinemia (NKH) is a rare inborn error of metabolism caused by impaired function of the glycine cleavage system (GCS) and characterised by accumulation of glycine in body fluids and tissues. NKH is an autosomal recessive condition and the majority of affected individuals carry mutations in GLDC (glycine decarboxylase). Current treatments for NKH have limited effect and are not curative. As a monogenic condition with known genetic causation, NKH is potentially amenable to gene therapy. An AAV9-based expression vector was designed to target sites of GCS activity. Using a ubiquitous promoter to drive expression of a GFP reporter, transduction of liver and brain was confirmed following intra-venous and/or intra-cerebroventricular administration to neonatal mice. Using the same capsid and promoter with transgenes to express mouse or human GLDC, vectors were then tested in GLDC-deficient mice that provide a model of NKH. GLDC-deficient mice exhibited elevated plasma glycine concentration and accumulation of glycine in liver and brain tissues as previously observed. Moreover, the folate profile indicated suppression of folate one‑carbon metabolism (FOCM) in brain tissue, as found at embryonic stages, and reduced abundance of FOCM metabolites including betaine and choline. Neonatal administration of vector achieved reinstatement of GLDC mRNA and protein expression in GLDC-deficient mice. Treated GLDC-deficient mice showed significant lowering of plasma glycine, confirming functionality of vector expressed protein. AAV9-GLDC treatment also led to lowering of brain tissue glycine, and normalisation of the folate profile indicating restoration of glycine-derived one‑carbon supply. These findings support the hypothesis that AAV-mediated gene therapy may offer potential in treatment of NKH.

Clinical laboratory techniques for detecting drugs of abuse: Feasibility in developing nations

Drugs of abuse are prescription, over-the-counter, or other forms of drugs that are often used for purposes other than those for which they are meant to be used, or in excessive amounts. The abuse of illicit drugs (for example cocaine, heroin, and codeine) poses a serious threat to public health, not to mention a great challenge to the health care system. It is important to review the techniques used in detecting these drugs of abuse to stay ahead of this growing global drug problem. This review aims to give an overview of known clinical laboratory techniques used in detecting some common drugs of abuse that could serve as an integral part of determining the presence of these drugs in bodily fluids and samples. Relevant literature was reviewed in various search engines (Google Scholar, PubMed, ScienceDirect, Bing). Findings showed that some of the current clinical laboratory techniques for detecting and quantifying drugs of abuse include Microcrystalline tests, thin-layer chromatography, colourimetric tests, immunoassays, urine dipstick tests, and ultraviolet spectroscopy. Apart from these known techniques, new and emerging techniques are being validated to serve as additions to these already existing techniques and be used in clinical settings. In conclusion, the burden of drug abuse is on the rise and becoming a public health concern. There should be an ever-increasing interest in developing new analytical methodologies not only to detect but also to quantify drugs of abuse which may be applied in a plethora of areas including clinical settings.Some of the outlined techniques are highly priced and for cost ineffectiveness, they may be difficult to afford and sustain in resource-restraint settings like developing nations. Hence, grants and laboratory infrastructural support may be needed from international donor agencies.

Bioabsorbable composite laminates of PLA reinforced with surface‐modified Mg wires for orthopedic implant applications

Unidirectional and multidirectional laminates of Mg wire‐reinforced PLA‐matrix composites were manufactured by an improved compression molding strategy that allows excellent control on the position and orientation of the wires. Two different types of Mg wires, with and without surface modification by continuous plasma electrolytic oxidation were used, the former to improve the interfacial strength and to reduce the degradation rate of Mg wires in biological environments. The mechanical properties of the constituents as well as of composites were measured before and after degradation by immersion in simulated body fluids and the corresponding deformation, fracture and degradation mechanisms were analyzed in detail. It was found that the presence of the Mg wires improved the mechanical behavior in tension and compression of unidirectional composites in the longitudinal direction (close to cortical bone) and that quasi‐isotropic laminates with tailored properties could be designed from the data of unidirectional composites.This article is protected by copyright. All rights reserved.

AZ31-calcium-deficient hydroxyapatite (CDHA) composites produced by friction stir processing: controlling the degradation by enhancing the biomineralisation

ABSTRACT In the present work, calcium deficient hydroxyapatite (CDHA) was dispersed into the surface of AZ31 Mg alloy within the solid state by friction stir processing and the role of the refined microstructure and the presence of nano-CDHA on biomineralization and corrosion performance was investigated. Fine grains (1.6 ± 2.1 µm) have been produced in AZ31-CDHA composite compared with base alloy (41 ± 6.1 µm) and nano-CDHA was observed as reinforced into AZ31 matrix. X-ray diffraction analysis confirms the texture change due to FSP in the composite. Wettability studies carried out by measuring contact angles on the surface of the composites indicated increased surface energy (37.27 ± 5.1 mJ/m2) compared with the base alloy (20.55 ± 3.4 mJ/m2) . From the potentiodynamic polarization studies, lower corrosion current density (1.69 ± 1.13 × 10-5 A/cm2) was observed for the composite. From the immersion studies in simulated body fluids, higher level of biomineralization on the composite helped to decrease the degradation rate as reflected from the lower weight loss (7.5 ± 1.1 %) compared with the base alloy (12.1 ± 1.3 %). The results demonstrate the promising role of enhanced biomineralization on controlling the degradation of AZ31-CDHA composite..

Identification of novel snoRNA-based biomarkers for clear cell renal cell carcinoma from urine-derived extracellular vesicles

BackgroundClear cell renal cell carcinoma (ccRCC) is the most common subtype of RCC with high rates of metastasis. Targeted therapies such as tyrosine kinase and checkpoint inhibitors have improved treatment success, but therapy-related side effects and tumor recurrence remain a challenge. As a result, ccRCC still have a high mortality rate. Early detection before metastasis has great potential to improve outcomes, but no suitable biomarker specific for ccRCC is available so far. Therefore, molecular biomarkers derived from body fluids have been investigated over the past decade. Among them, RNAs from urine-derived extracellular vesicles (EVs) are very promising.MethodsRNA was extracted from urine-derived EVs from a cohort of 78 subjects (54 ccRCC patients, 24 urolithiasis controls). RNA-seq was performed on the discovery cohort, a subset of the whole cohort (47 ccRCC, 16 urolithiasis). Reads were then mapped to the genome, and expression was quantified based on 100 nt long contiguous genomic regions. Cluster analysis and differential region expression analysis were performed with adjustment for age and gender. The candidate biomarkers were validated by qPCR in the entire cohort. Receiver operating characteristic, area under the curve and odds ratios were used to evaluate the diagnostic potential of the models.ResultsAn initial cluster analysis of RNA-seq expression data showed separation by the subjects’ gender, but not by tumor status. Therefore, the following analyses were done, adjusting for gender and age. The regions differentially expressed between ccRCC and urolithiasis patients mainly overlapped with small nucleolar RNAs (snoRNAs). The differential expression of four snoRNAs (SNORD99, SNORD22, SNORD26, SNORA50C) was validated by quantitative PCR. Confounder-adjusted regression models were then used to classify the validation cohort into ccRCC and tumor-free subjects. Corresponding accuracies ranged from 0.654 to 0.744. Models combining multiple genes and the risk factors obesity and hypertension showed improved diagnostic performance with an accuracy of up to 0.811 for SNORD99 and SNORA50C (p = 0.0091).ConclusionsOur study uncovered four previously unrecognized snoRNA biomarkers from urine-derived EVs, advancing the search for a robust, easy-to-use ccRCC screening method.

Chromatographic methods and approaches for bioequivalence study, drug screening and enantioseparation of indapamide

AbstractIndapamide (Indp) and certain other diuretics have been abused in sports, therefore, having sensitive methods for its detection and assay in biological fluids (whole blood, plasma, serum, and urine) is of significant importance. The racemic mixture of Indp is being used as an active pharmaceutical ingredient among other commonly prescribed diuretics. The regulatory authorities and pharmaceutical industries demand analytical methods for successful enantioseparation of such molecules. The paper presents a critical overview of the scientific issues of the application of contemporary techniques involving various chromatographic approaches (with liquid or supercritical fluid as mobile phases) and capillary electrophoresis and method development, for drug screening, assay, bioequivalence studies and enantioseparation of indapamide with their results. It also covers the historical developments that led to significant breakthroughs in research and concise evaluations of research in the area.Different types of chromatographic methods (HPLC, CEC, SFC etc) discussed herein provide an insight and a choice to select a method to (i) screen Indp for drug abuse, (ii) separate, isolate and quantify the enantiomers of Indp and (iii) investigate their pharmacokinetics as markedly different species and not as a total drug. The article evaluates the field's status with a broad base and practical oriented approach so that the underlying principles are easily understood to help chemists and non-specialists gain useful insights into the field outside their specialization and provide experts with summaries of key developments. To the best of authors' knowledge there has been no attempt to review such methods for analysis of Indp and this is the first report of its kind.