Analysis Of Body Fluids Research Articles

Determination of multiple analytes in urine using label-free SERS coupled with simple sample pretreatments

BackgroundA key restriction of label-free surface-enhanced Raman spectroscopy (SERS) in analysis of objects with complex composition (including with several target analytes) is the competition of mixture components for interaction with SERS-active surface. This leads to poor selectivity of the analysis of such mixtures (e.g., body fluids) and the need to use advanced sample pretreatment procedures such as HPLC or TLC. Therefore, this work aims to develop a set of simple and fast pretreatment steps (dilution, pH correction, etc.) to increase the sorption of the target analyte, reduce the sorption of admixtures, and prevent suppression of the target analyte SERS signal. ResultsWe have developed label-free SERS assay suitable for the determination of three analytes (methotrexate, cephalosporin antibiotic, and creatinine) in one real urine sample as a model matrix with complex and deviating composition. The choice of drugs is justified by the need to monitor their concentration in urine during joint drug treatment of cancer patients with concomitant bacterial infection, while monitoring creatinine concentration helps to evaluate kidney function of the patients. Additionally, three cephalosporin representatives were used in the study to maximize versatility of the assay. As a results, the optimized pretreatment steps enable to eliminate the negative influence of excess of interferences (including other analytes) and achieve precise (≤12 % RSD) and accurate (88–111 % recovery) determination of several analytes in the therapeutically relevant ranges: 300–3000 μg mL−1 for creatinine, 20–200 μg mL−1 for methotrexate and cephalosporins. SignificanceTherefore, in addition to reporting a new SERS assay for the analysis of body fluids, this study clearly demonstrates the importance of taking into account competitive adsorption processes on the SERS substrate surface. We suggest making this practice mandatory when developing any label-free SERS assay because it enables to maximize the selectivity and accuracy of the analysis as well as to simplify the analysis procedure.

A Nonfouling Electrochemical Biosensor for Protein Analysis in Complex Body Fluids Based on Multifunctional Peptide Conjugated with PEGlyated Phospholipid.

Developing antifouling biosensors capable of performing robustly in complex human body fluids is crucial for biomarker diagnosis and health monitoring. Herein, an antifouling and highly sensitive and stable biosensor was constructed through the self-assembly of the designed conjugates composed of a multifunctional peptide (MP) and PEGylated distearoylphosphatidylethanolamine (DSPE-PEG). The self-assembly capability of the DSPE-PEG-MP was demonstrated clearly through coarse-grained molecular dynamics simulation and transmission electron microscopy, and it can be effectively self-assembled onto the electrode surface modified with gold nanoparticles. The MP was designed to be antifouling and contained a peptide sequence that can specifically bind the target protein Annexin A1 (ANXA1), and the D-type amino acid composition of MP can enhance its resistance to enzymatic hydrolysis. The unique design of MP, in conjugation with the self-assembly capability of the PEGylated phospholipid DSPE-PEG, enabled the biosensor to exhibit excellent antifouling capability and stability in various complex human body fluids. The biosensor was capable of sensitively and selectively quantifying ANXA1 and achieved a limit of detection down to 0.12 pg mL-1. More importantly, the biosensor demonstrated satisfactory accuracy for ANXA1 detection in clinical serum samples, as verified by the enzyme linked immunosorbent assay (ELISA) kits. It is expected that various antifouling biosensors suitable for application in complex biological environments can be constructed by utilizing the strategy of designing similar DSPE-PEG-MP conjugates.

Study of Co-relation between Clinical and Diagnostic Profile of Pediatric Tuberculosis in a Tertiary Care Hospital

Objective: Study of Co-relation between Clinical and Diagnostic profile of Pediatric Tuberculosis in a tertiary care hospital. Methods: This hospital-based observational study was conducted at the Department of Pediatrics, Muzaffarnagar Medical College and Hospital, covering a population of children aged <18 years diagnosed with TB. Data collection spanned 18 months, with a sample size of 100 children. Results: The study identified a predominant TB incidence in children aged 11-17 years (73%) with more females affected (66%) than males. Common symptoms included fever, cough, weight loss, and loss of appetite. Diagnostic findings showed a high use of Chest X-rays and Molecular tests. PTB was the most common form, accounting for 47 cases, EPTB accounting for 48 cases and Disseminated form was seen in 5 cases. Conclusions: Chest X-ray, Tuberculin Skin Test, Molecular Tests, and Blood Tests were universally performed on all patients. Microbiological confirmation is more common in pulmonary tuberculosis cases, while radiology, FNAC/Biopsy, and body fluid analysis are more utilized for extrapulmonary/disseminated cases. Diagnosing tuberculosis (TB) in children is often challenging due to several factors, including the difficulty in obtaining specimens and the typical paucibacillary nature.

A-158 Multi-site verification and performance of body fluids in-between Horiba Medical Yumizen h2500 with Sysmex XN-10 automated hematology analyzers and manual microscopy for morphology assessment

Abstract Background The analysis of body fluids plays an important role in the diagnosis and management of a wide variety of conditions. As standards for method validation and laboratory developed tests evolve, the body fluids analysis is receiving increased attention from both laboratories and regulatory bodies. We present the evaluated result of the validation and clinical performance accuracy of HORIBA Medical Yumizen H2500 analyzers as compared to SYSMEX XN-10 analyzers and to manual slide microscopy examination as a reference method for differential analysis for body fluids samples. Methods Depending on each laboratory collected procedures, synovial samples were collected with Sodium Heparin or K2EDTA or without anticoagulant and the other samples were collected with K2EDTA or without anticoagulant. As recommended, most (94%) of the synovial fluids were treated with the hyaluronidase. For the comparison of Yumizen H2500 versus XN-10 analyzers, the study was performed in 2 US sites and 1 French site collecting 390 samples totally: 138 serous fluids (Ascites, Pleural, Peritoneal, Pericardial), 169 synovial fluids and 83 CSF fluids. Samples were analyzed as soon as possible after collection (with a limit of 24 hours for serous and synovial fluids and 4 hours for CSF) and within 2 hours between both instruments. For the comparison of Yumizen H2500 versus reference method (manual microscopy), the study was performed in 2 US sites collecting 58 samples: 34 serous fluids (Ascites, Pleural, Peritoneal, Pericardial), 12 synovial fluids and 12 CSF fluids. Two qualified morphology examiners made the manual microscopy analysis and the average of the results of the 2 slide readings was used for the analysis. Results Considering all types of body fluids, the H2500/XN results are very well correlated for BFWBC (0.969), BFRBC (0.999), BFMN% (0.912), BFMN# (0.89), BFPN% (0.912), BFPN# (0.97). Similarly, the statistical analysis per body fluids type shows very good correlation. For serous type, all results are higher than 0.876, for synovial type all results are higher than 0.879 and for cerebrospinal fluids type, all results are higher than 0.788. Among all samples, less than 8 samples (<2%) were out of the 95% interval. The comparison Yumizen H2500 versus manual microscopy, all the correlation results (r²) are very good: 0,9705 for Polynuclear and 0,9742 for Mononuclear. The study demonstrated better results of Yumizen H2500 versus manual microscopy than versus XN-10 for BFPN% (0.9705 versus 0.912) and BFMN% (0.9742 versus 0.912). Among 58 samples, only 4 (6.9%) were out of the 95% interval for BFPN% and BFMN%. Conclusions The Yumizen H2500 results for body fluid WBC differentiation (Polynuclear and Mononuclear) are very well correlated with the results from manual slide microscopy evaluation. The comparison of the results analyzed on Yumizen H2500 and on XN does not show any significant difference.

A-154 Non-standard body fluid validation of d-dimer analysis of organox liver perfusate to support clinical evaluations of donor livers prior to transplantation

Abstract Background Non-standard body fluid analysis is considered a challenge to laboratories since very few have been approved by the FDA for routine testing. As medical procedures evolve, the laboratory must balance the requirement to uphold regulatory standards while also supporting procedures that improve patient care. The LUMC Liver Transplant Team recently implemented the use of the FDA-approved OrganOx Metra normothermic machine perfusion instrument for maintaining donor livers under physiologic conditions prior to transplantation. The team mitigates risk for biliary complications associated with fibrin microthrombi by perfusing a combination of alteplase rTPA with fresh frozen plasma and monitoring the prevalence of fibrin through the release of D-dimers over the course of ex situ normothermic preservation. Liver perfusate is not an FDA-approved specimen type for the analysis of D-dimer and requires laboratory verification as a compatible specimen for analysis. Methods The D-dimer HS 500 assay was performed on an ACL TOP 750 analyzer. Carry-over studies evaluated activated partial thromboplastin time (aPTT) of a normal control (citrated plasma) specimen initially, and repeatedly in-between sets of D-dimer measurements of heparinized perfusate specimen. Recovery studies were performed by mixing liver perfusate with citrated plasma spiked with D-dimer in known ratios, allowing for the calculation of an expected value of D-dimer. Results Carry-over studies measuring aPTTs of normal plasma specimens provided %CVs of 1 and 3%. Recovery of D-dimer in seven sets of independent add-mixture series (three mixed specimen per set) was on average 105% with a range from 95 to 113% recovery. Conclusions Concern for cross-contamination of perfusate specimen containing heparin anticoagulant on analyzers used to evaluate routine coagulation testing was mitigated by the results of the carry-over study, which provided %CV within the manufacturer limit for intraday precision. Recovery studies evaluating matrix effect for the analysis of D-dimer on liver perfusate generated under the LUMC liver transplant protocol indicated that the matrix of the perfusate did not impact D-dimer analysis, as compared to the FDA-approved specimen type.

Near‐Infrared Spectroscopy and Aquaphotomics in Cancer Research: A Pilot Study

ABSTRACTCurrently, the majority of methods to monitor cancer treatment through the analysis of body fluids are based on a highly selective detection of single molecules or cells. In this study, we are considering the analysis of the aqueous medium of liquid samples, that is, water, itself, using aquaphotomics and near‐infrared spectroscopy (NIR) for spectral data acquisition and processing, within cancer research. Water, as a molecular system, is a rich source of information about the current state of a patient, which can be extracted from near‐infrared spectra of liquid samples via simple algorithms based on multivariate data analysis. The reported results, obtained ex vivo of body fluids, demonstrate the potential of aquaphotomics in cancer research.

Untargeted Metabolomic Analyses of Body Fluids to Differentiate TBI DOC and NTBI DOC.

To investigate the metabolomic differences between Traumatic brain injury (TBI) disorder of consciousness (DOC) patients and non-traumatic brain injury (NTBI) DOC patients by using cerebrospinal fluid (CSF), serum and urine samples beneficial to understand the pathological mechanism differences between the two etiologies, provide potential clues for the subsequent treatment and prognosis, and investigate the metabolome differences and similarities between TBI and NTBI among three different body fluids. In total, 24 TBI DOC subjects and 29 NTBI DOC subjects were enrolled. CSF, serum and urine samples from TBI DOC and NTBI DOC patients were collected and analyzed by performing UPLC-MS. The statistical methods and pathway analyses were applied to discover potential biomarkers and altered metabolic functions. When comparing TBI DOC and NTBI DOC, 36, 31 and 52 differential metabolites were obtained in CSF, serum and urine, respectively. The functional analysis of differential metabolites obtained in CSF, serum and urine were all related to amino acid metabolism. Except for amino acid metabolism, metabolic biomarkers in CSF, serum and urine mainly focus on central function, cognitive function, necrosis and apoptosis and neurological function, respectively. In CSF, the highest AUC was 0.864 (Isoproturon) and 0.816 (Proline betaine). Then, the AUC of NFurfurylformamide in serum was 0.941, while the AUC of Dihydronepetalactone and Doxepin N-oxide glucuronide were 1.0 in urine. CSF, serum and urine metabolomic analyses could differentiate TBI DOC from NTBI DOC and functional analyses showed a metabolic change difference between TBI DOC and NTBI DOC.

Identification of blood at simulated crime scenes using silver nanoparticles with SERS.

The analysis of substances and samples obtained from a crime scene is very important in solving forensic cases. To determine the variables involved in a crime and to expedite the investigation process, the rapid analysis of body fluids in small quantities and within environments containing diverse components is particularly necessary. For this reason, it is of great importance to analyze biological fluids with rapid, noncontaminating, nondestructive, low-cost, and accurate techniques. In recent years, with advancements in laser technology, spectroscopic methods have been introduced as analytical techniques in forensic medicine and chemical studies. This study focuses on surface-enhanced Raman spectroscopy (SERS) to demonstrate the detection of blood samples in simulated crime scenes. To minimize the background signal from fluorescent biomolecules in blood, dilution was performed with two different components and Raman analysis was performed for four different concentrations of blood. In general, a decrease in noise in the spectra was observed as the blood was diluted. Crime scenes consisting of pure blood, blood diluted with ethanol and distilled water (1:2, 1:4, and 1:8), a blood-mineral water mixture, a blood-cherry juice mixture, and silver nanoparticle-added mixtures were simulated, and their spectra were examined. Chemometric analyses of the data were performed. Despite high noise and low peak intensities, blood-identifying signals were detected when examining different blood concentrations. It was observed that silver nanoparticles provided high enhancement of blood peaks thanks to their strong plasmonic properties.

Antifouling and antimicrobial wearable electrochemical sweat sensors for accurate dopamine monitoring based on amyloid albumin composite hydrogels

Wearable electrochemical sweat sensors are potentially promising for health monitoring in a continuous and non-invasive mode with high sensitivity. However, due to the complexity of sweat composition and the growth of skin bacteria, the wearable sweat sensors may gradually lose their sensitivity or even fail over time. To deal with this issue, herein, we proposed a new strategy to construct wearable sweat sensors with antifouling and antimicrobial capabilities. Amyloid albumin hydrogels (ABSAG) were doped with two-dimensional (2D) nanomaterial MXene and CeO2 nanorods to obtain the antifouling and antimicrobial amyloid albumin composite hydrogels (ABSACG, CeO2/MXene/ABSAG), and the wearable sensors were prepared by modifying flexible screen-printed electrodes with the ABSACG. Within this sensing system, the hydrophilic ABSAG possesses strong hydration capability, and it can form a hydration layer on the electrode surface to resist biofouling in sweat. The 2D nanomaterial MXene dispersed in the hydrogel endows the hydrogel with good conductivity and electrocatalytic capability, while the doping of CeO2 nanorods further improves the electrocatalytic performance of the hydrogel and also provides excellent antimicrobial capability. The designed wearable electrochemical sensors based on the ABSACG demonstrated satisfying antifouling and antimicrobial abilities, and they were capable of detecting dopamine accurately in human sweat. It is expected that wearable sensors utilizing the antifouling and antimicrobial ABSACG may find practical applications in human body fluids analysis and health monitoring.

Ethanol concentrations in various biological specimens: Living and postmortem forensic toxicology analysis and comprehensive literature review

Alcohol use upsurges the risk for many chronic ill-health consequences such as hepatitis, malignancies, and disastrous outcomes like road traffic accidents ending in fatal injuries. Biochemical and toxicological analysis of different body fluids is crucial for identifying the cause of death and postmortem interval in many forensic cases. Blood, urine, and vitreous fluid are the most valuable body fluids for detecting alcohol during any toxicological analysis. Alcohol is responsible for widespread morbidity and mortality worldwide. Blood alcohol concentration (BAC) is a necessary toxicological test to investigate various crime and accident scenes. This study comprehensively explores the demographic characteristics, BAC distribution, and correlations of alcohol concentrations in postmortem and living cases. Postmortem cases (N = 166) reveal intriguing demographic patterns, with notable variations in year distribution, nationality, sex, age groups, occupation, smoking habits, place of death, and psychiatric history. Living cases (N = 483) exhibit distinct demographic profiles, emphasizing differences in year distribution, nationality, sex, age groups, and smoking habits.Analysis of BAC distribution reveals diverse patterns in both postmortem and living cases, providing valuable insights into the prevalence of different BAC levels in each group. Correlation analyses unveil strong associations between alcohol concentrations in various biological samples in postmortem cases, highlighting the interdependence of blood, vitreous, and urine alcohol concentrations. Conversely, living cases display a moderate positive correlation between blood and urine alcohol concentrations.Comparative analyses showcase significant differences in mean alcohol concentrations between postmortem and living cases, suggesting variations in alcohol metabolism and distribution. These findings underscore the importance of considering temporal factors in interpreting alcohol concentrations in forensic and clinical contexts.In conclusion, this study enhances our understanding of alcohol-related incidents by delineating demographic profiles, BAC distributions, and correlations between different biological samples. Such insights are crucial for refining investigative and clinical approaches, contributing to the broader fields of forensic science and public health.

Determination of ABO blood group based on secretors or non-secretors analysis in body fluids

Criminal acts often leave traces analyzed by law enforcement, especially by the Indonesian National Police (POLRI). Sometimes, various items were found and analyzed to link the victim and the perpetrator. Blood, the most abundant body fluid, found in crime scenes, consists of cellular and liquid elements. The forensic focuses on red blood, which can be wet or dried at the crime scene. Sometimes, there was no blood or bloodstains found, but objects related to other body fluids, such as: condoms, toothbrushes, the mouth of glass cups, straws, etc, could be found. Blood group analysis could still be carried out based on secretor gene (Se) considering that 80% of the population was of the secretor group. There are two chromosomes related to blood regulation, namely: chromosomes no. 9 and 19. The former relates to the ABO blood group and the latter relates to the secretory phenotype. The basic principle of secretors was antigens found on the surface of red blood cells and all body fluids, except cerebrospinal fluid. The pattern of inheritance on chromosomes 9 and 19 is based on the probability law by Mendell. It stated that all of genotypes (homozygous dominant-recessive or heterozygous) would be passed on to their offspring.

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.

Potential application of body fluids autofluorescence in the non-invasive diagnosis of endometrial cancer.

Endometrial carcinoma (EC) is the most common cancer of the female reproductive tract in developed countries. The prognosis and 5-year survival rates are closely tied to the stage diagnosis. Current routine diagnostic methods of EC are either lacking specificity or are uncomfortable, invasive and painful for the patient. As of now, the gold diagnostic standard is endometrial biopsy. Early and non-invasive diagnosis of EC requires the identification of new biomarkers of disease and a screening test applicable to routine laboratory diagnostics. The application of untargeted metabolomics combined with artificial intelligence and biostatistics tools has the potential to qualitatively and quantitatively represent the metabolome, but its introduction into routine diagnostics is currently unrealistic due to the financial, time and interpretation challenges. Fluorescence spectral analysis of body fluids utilizes autofluorescence of certain metabolites to define the composition of the metabolome under physiological conditions. This review highlights the potential of fluorescence spectroscopy in the early detection of EC. Data obtained by three-dimensional fluorescence spectroscopy define the quantitative and qualitative composition of the complex fluorescent metabolome and are useful for identifying biochemical metabolic changes associated with endometrial carcinogenesis. Autofluorescence of biological fluids has the prospect of providing new molecular markers of EC. By integrating machine learning and artificial intelligence algorithms in the data analysis of the fluorescent metabolome, this technique has great potential to be implemented in routine laboratory diagnostics.

Wide-Targeted Semi-Quantitative Analysis of Acidic Glycosphingolipids in Cell Lines and Urine to Develop Potential Screening Biomarkers for Renal Cell Carcinoma.

Glycosphingolipids (GSLs), mainly located in the cell membrane, play various roles in cancer cell function. GSLs have potential as renal cell carcinoma (RCC) biomarkers; however, their analysis in body fluids is challenging because of the complexity of numerous glycans and ceramides. Therefore, we applied wide-targeted lipidomics using liquid chromatography-tandem mass spectrometry (LC-MS/MS) with selected reaction monitoring (SRM) based on theoretical mass to perform a comprehensive measurement of GSLs and evaluate their potency as urinary biomarkers. In semi-quantitative lipidomics, 240 SRM transitions were set based on the reported/speculated structures. We verified the feasibility of measuring GSLs in cells and medium and found that disialosyl globopentaosylceramide (DSGb5 (d18:1/16:0)) increased GSL in the ACHN medium. LC-MS/MS analysis of urine samples from clear cell RCC (ccRCC) patients and healthy controls showed a significant increase in the peak intensity of urinary DSGb5 (d18:1/16:0) in the ccRCC group compared with that in the control group. Receiver operating characteristic analysis indicated that urinary DSGb5 could serve as a sensitive and specific marker for RCC screening, with an AUC of 0.89. This study demonstrated the possibility of urinary screening using DSGb5 (d18:1/16:0). In conclusion, urinary DSGb5 (d18:1/16:0) was a potential biomarker for cancer screening, which could contribute to the treatment of RCC patients.

Application of the International System for Reporting Serous Fluid Cytopathology: An Experience at a Tertiary Care Centre in India

Abstract Background: The cytological analysis of body fluids is the preliminary investigation performed for the diagnostic evaluation of body fluids, helping the physician make the diagnosis and plan proper treatment. The International System for Reporting Serous Fluid Cytology (ISFRSFC) was created using best international practises, current literature and expert consensus to make a uniform reporting system for serous fluids. Aims: The present study was conducted with the aim of recategorising the effusion fluids as per the guidelines prescribed by ISRFRC, knowing the distribution of cases into various categories and assessing their feasibility in day-to-day reporting. Patients and Methods: This study was performed on serous fluids (ascitic, pleural, peritoneal and pericardial) received over a period of 5 years, from January 2017 to April 2022, in the cytology unit of the Department of Pathology, ABVIMS, Dr. Ram Manohar Lohia Hospital, New Delhi. Wherever possible, clinicoradiological and histopathological data were obtained and correlated with cytological findings. Serous fluids were categorised into reporting formats as prescribed by ISRFRC and its feasibility was assessed in day-to-day reporting. Results: A total of 1115 cases of serous effusion fluid were examined. The male-to-female ratio was 1:1.12. Pleural fluid constituted the majority of the sample (436, 39.10%). Overall, 56 (5.02%) cases were non-diagnostic (ND), 1013 (90.85%) cases were negative for malignancy, 8 (0.72%) were atypia of undetermined significance, 11 (0.99%) were suspicious of malignancy and 27 (2.42%) were malignant (MAL). Conclusion: The current study emphasises the significance of cytological analysis of serous fluid as a comprehensive diagnostic investigation that also aids in determining the cause and monitoring the disease process. With the introduction of ISFRSFC, serous fluid reporting is now uniform and more comprehensive, both for cytopathologists and clinicians.

A case of tuberculous peritonitis accompanied by tuberculous pleuritis

Tuberculous peritonitis is a form of extrapulmonary tuberculosis, a peritoneal or visceral inflammation caused by Mycobacterium tuberculosis. The disease is rarely independent, but is usually a continuation of the tuberculosis process elsewhere, especially pulmonary tuberculosis. We report a case of TB peritonitis accompanied by TB pleurisy, a 29-year-old female patient with complaints of an enlarged abdomen, heartburn, fever, diarrhea, and decreased appetite. Treatment history was Acitral, Zinc, and Metronidazole. On physical examination, it was found that the general condition was weak and the axilla temperature was 39.5ºC. Thorax examination; decreased vesicular sound on the left chest. Abdominal examination; found distension, epigastric tenderness, undulation, shifting dullness, checkerboard phenomenon, and increased bowel noise. Laboratory examination of complete blood; within normal limits. Complete stool; yellow color, mucus (+), leukocytes 4-6/LPB, bacteria (+). The thorax photo showed left pleural effusion, BOF 3 position: ascites. Abdominal ultrasound results: thickening of the peritoneum, ascites, suspected TB peritonitis. Results of ascites and pleural fluid analysis: rivalta (+) and Adenosine Deaminase (ADA) increased. So that from anamnesis, physical examination, supporting examination can be established diagnosis of TB Peritonitis and TB Pleuritis, followed by OAT therapy; FDC for 12 months. From this case it can be concluded that clinical and supporting examinations (radiology) are needed to make a correct diagnosis and body fluid analysis examinations can help confirm the diagnosis.

Surface‐Enhanced Raman Spectroscopy‐Based Optical Biosensor for Liquid Biopsy: Toward Precision Medicine

AbstractLiquid biopsy is regarded as a promising strategy for assisting precision medicine because of its convenience, noninvasiveness, and ability to overcome tumor heterogeneity and achieve early detection. Recently, impressive advancements in plasmonic biosensors, artificial intelligence, and portable Raman equipment have yielded unprecedented progress in surface‐enhanced Raman spectroscopy (SERS)–based point‐of‐care testing (POCT) systems for liquid biopsy. The development of these systems presents a paradigm shift in on‐site liquid biopsy applications by leveraging the unique benefits of efficiency, fast analysis, portability, affordability, and user‐friendliness. Herein, these advances are introduced over the last 3 years in the field of SERS‐based POCT systems for labeled and label‐free biomarker analysis in body fluids, including tumor circulating proteins and cells, exosomes, micro‐RNA, and circulating tumor DNA. Additionally, powerful machine learning algorithms (including deep learning algorithms) are integrated with SERS to effectively extract potential data features and generate precise diagnostic models. The review highlights the use of handheld and portable Raman devices in significantly promoting the application of SERS‐based POCT in clinical scenarios. Finally, the review outlines the challenges and future perspectives of this technology.

Validation of N-Methylpyridinium as a Feasible Biomarker for Roasted Coffee Intake

Health-related nutritional human studies rely on the validity of dietary data provided by study participants. Reliable biomarkers for food intake help objectify data collected by food frequency questionnaires. They facilitate the monitoring of compliance with the study requirements, e.g., abstinence from food, help clean biased data, and remove non-compliant individuals. Biomarker candidates are often revealed by sophisticated metabolomics analyses of body fluids, e.g., urine or plasma, collected from case and control study populations. However, validation for using a biomarker candidate in real-life scenarios is seldomly executed. Coffee is a food item of high interest because of the abundance of bioactive compounds and the regularity of life-time consumption by a large part of the population. Coffee has been found to positively impact cardiovascular risk, type 2 diabetes, and cognitive decline. Coffee and its health implications, therefore, are of high interest. A suitable dietary biomarker for coffee consumption is desirable for the clear classification of study participants as coffee drinkers or non-coffee drinkers to enable correlation of physiological response to dietary habits, e.g., coffee consumption. Here, we propose the roast coffee compound N-methylpyridinium (NMP) as a promising biomarker of pragmatic use to distinguish a coffee drinker from a non-coffee drinker. NMP is an easily accessible analytical target from the plasma and urine matrix that can help determine precedent exposure to roasted coffee products. We review the published information on the coffee compound N-methylpyridinium in foods, coffee, and plasma/urine after coffee consumption, and evaluate the data in the context of the proposed food biomarker criteria “plausibility”, “time- and dose–response”, “robustness”, “reliability”, “stability”, “analytical performance”, and “reproducibility”. An additional data set is acquired to fill the gaps in the literature. In summary, we conclude that the abundance of NMP can serve as a reliable analytical tool to verify recent consumption of roasted coffee. The use of NMP appears limited to being qualitative, as NMP abundance in coffee and human biosamples is affected by several parameters, e.g., the roasting conditions and the volume and time of coffee consumed.

Ciljanje onkogenih puteva posredstvom laktoferina u hemioprevenciji raka i dopunskoj terapiji - od mehanističkih uvida do kliničkih ispitivanja

Genetic, genomic and proteomic analyses of cells, tissues and body fluids have generated a wealth of precious information about the intricate mechanisms which underlie carcinogenesis and metastasis. Lactoferrin, a multifunctional cationic glycoprotein, has attracted widespread appreciation because of its characteristically novel properties for cancer chemoprevention. Tumor microenvironment is a highly complicated and sophisticated ecosystem, significantly reshaped by a wide variety of treatment regimes. Therefore, lactoferrin-mediated immunostimulatory role reshapes tumor microenvironment and inhibits cancer progression. There is sufficient experimental evidence related to immunostimulatory ability of lactoferrin in tumor microenvironment. Different clinical trials have been conducted for the evaluation of clinical efficacy of lactoferrin in different cancer patients. It is necessary to carefully interpret the clinical evidence and identify the major gaps in our understanding related to the selection of group of cancer patients likely to benefit the most from the combinatorial treatment regime comprised of lactoferrin and chemotherapeutic drugs. Moreover, lack of efficacy should be analyzed by a team of interdisciplinary researchers for a broader and comprehensive understanding of the mechanisms underlying treatment failure.

Engineering a universal and fully automatic analyzer for multichannel and on-site biochemical detection in body fluids.

Rising concerns over wellness and aging have heightened the demand for convenient and efficient on-site health monitoring and disease screening. Current research, focused on specific biomarker detection, often neglects the complexities of sample matrix interference and the absence of a comprehensive, automated platform. To address these issues, we have developed a universal, fully automated analyzer for multifaceted, on-site biochemical analysis of body fluids. This analyzer integrates automated sample pretreatment, automatic dilution, detection, and self-cleaning functionalities seamlessly. It is designed to detect a wide range of analytes, from small molecules to macromolecules, including ions and proteins, utilizing spectrophotometric sensing. After optimization, the analyzer achieves performance comparable to traditional Enzyme-Linked Immunosorbent Assay (ELISA), while significantly expanding its detection range through automated dilution. Demonstrations of small molecule detection include the simultaneous assessment of citric acid (CA) and oxalic acid (OA) in urine, achieving recovery rates between 96.65%-106.42% and 93.13%-112.50%, respectively. For protein detection, the analyzer successfully identified Cyfra21-1 in saliva with a recovery rate of 104.93%-111.31%. The pre-treatment process requires only 8.8 minutes, showing enhanced recovery rates for CA and OA at 97.8% and 97.6% respectively, which are superior and more rapid than manual methods. The exemplary pretreatment and detection performance of the analyzer underlines its effectiveness in multifaceted, on-site biomarker detection, establishing it as a promising and versatile tool for disease screening and health monitoring. This analyzer offers a powerful technological solution for on-site fluid testing, advancing community health care by facilitating more reliable and rapid diagnostics.