Body Fluid Samples Research Articles

Metabolomic signatures associated with fetal growth restriction and small for gestational age: a systematic review

AbstractThe pathways involved in the pathophysiology of fetal growth restriction (FGR) and small for gestational age (SGA) are incompletely understood. We conduct a systematic review to identify metabolomic signatures in maternal and newborn tissues and body fluids samples associated with FGR/SGA. Here, we report that 825 non-duplicated metabolites were significantly altered across the 48 included studies using 10 different human biological samples, of which only 56 (17 amino acids, 12 acylcarnitines, 11 glycerophosphocholines, six fatty acids, two hydroxy acids, and eight other metabolites) were significantly and consistently up- or down-regulated in more than one study. Three amino acid metabolism-related pathways and one related with lipid metabolism are significantly associated with FGR and/or SGA: biosynthesis of unsaturated fatty acids in umbilical cord blood, and phenylalanine, tyrosine and tryptophan biosynthesis, valine, leucine and isoleucine biosynthesis, and phenylalanine metabolism in newborn dried blood spot. Significantly enriched metabolic pathways were not identified in the remaining biological samples. Whether these metabolites are in the causal pathways or are biomarkers of fetal nutritional deficiency needs to be explored in large, well-phenotyped cohorts.

Bacterial profile-based body fluid identification using a machine learning approach.

Identifying the origins of biological traces is critical for the reconstruction of crime scenes in forensic investigations. Traditional methods for body fluid identification rely on chemical, enzymatic, immunological, and spectroscopic techniques, which can be sample-consuming and depend on simple color-change reactions. However, these methods have limitations when residual samples are insufficient after DNA extraction. This study aimed to develop a method for body fluid identification by leveraging bacterial DNA profiling to overcome the limitations of the conventional approaches. Bacterial profiles were determined by sequencing the hypervariable region of the 16S rRNA gene,using DNA metabarcoding of evidence collected from criminal cases. Amplicon sequence variants (ASVs) were analyzed to identify significant microbial patterns in different body fluid samples. The bacterial profile-based method demonstrated high discriminatory power with a machine learning model trained using the naïve Bayes algorithm, achieving an accuracy of over 98% in classifying samples into one of four body fluid types: blood, saliva, vaginal secretion, and mixture traces of vaginal secretions and semen. Bacterial profiling enhances the accuracy and robustness of body fluid identification in forensic analysis, providing a valuable alternative to traditional methods by utilizing DNA and microbial community data despite the uncontrollable conditions. This approach offers significant improvements in the classification accuracy and practical applicability in forensic investigations.

Environmental microbiota from substrate may interfere with microbiome-based identification of forensically relevant body fluids: A pilot study

The microbiome is a promising tool for identifying body fluids which can be deposited on various substrates at a crime scene. Body fluids collected from crime scenes are not entirely free from substrate microbes whose effects on the microbiome-based identification of body fluids are not well understood. In this study, five body fluids (peripheral blood, menstrual blood, nasal secretions, saliva, and semen) were deposited on sterile swabs, bedspreads, and floors under indoor exposure conditions for 7 days. The microbial communities in the samples were characterized using amplicon sequencing targeted V4 region of 16S rRNA gene. The results showed that the microbial communities of fresh samples deposited on sterile swabs clustered together according to the type of body fluid. The microbial composition of the body fluids deposited on the bedspread and floor is significant different from those deposited on sterile swabs. The microbial communities of mock body fluids were a mixture of microbes from pure body fluids and environmental microbes. FEAST analysis showed that the microbes of mock saliva samples were mainly from pure body fluids (51.53 % and 63.04 % on the bedspread and floor, respectively), but not from substrates (25.70 % and 18.92 % on the bedspread and floor, respectively). Contrary results were observed in peripheral blood, mock nasal secretion, and semen samples. All samples were mainly clustered based on the substrate, but not on the type of body fluid in the PCoA visualization. PERMANOVA results showed that the substrate accounted for more of the variance (R2 = 0.211, P < 0.001) than the type of body fluid (R2 = 0.152, P < 0.001). MicroDecon was used to remove contamination by microbes from the substrate of mock body fluid samples. PCoA and PERMANOVA were performed using decontaminated data. The results showed that samples were no longer clustered based on the substrate, and the type of body fluid (R2 = 0.240, P < 0.001) accounted for more of the variance in the microbial communities of samples than the substrate (R2 = 0.108, P < 0.001). Our results suggest that environmental microbiota from substrates may interfere with the microbiome-based identification of forensically relevant body fluids. To some extent, decontamination could decrease the effects of the substrate on the microbial communities of the samples and enhance the ability to distinguish between the types of body fluids. This pilot study will be valuable in promoting the application of microbiome-based stain analysis in forensics.

Comparison of DNA extraction procedures for detection of Mycoplasma bovis directly from extended bovine semen straw samples using a commercial M. bovis PCR

BackgroundMycoplasma bovis is a global pathogen of cattle but was detected for the first time in New Zealand in 2017, triggering a response under their Biosecurity Act as an “unwanted organism”. Following a lengthy eradication campaign, the Ministry of Primary Industries (MPI) now requires all bovine semen destined for export to New Zealand to be screened with an M. bovis-specific real-time PCR (rtPCR) compliant with amended import health standard (IHS) test requirements aimed at preventing the accidental importation of M. bovis. The standard stipulates that semen samples cannot be centrifuged prior to DNA extraction. To comply with these strict requirements, one of the listed tests was validated together with different DNA preparation steps and compared with existing in-house procedures. DNA was extracted from semen straws using the current in-house semi-automated platform procedures for processing culture, tissue and body fluid sample submissions and was compared with the stipulated test requirements. DNA from centrifuged and unspun semen samples spiked with M. bovis was also compared.ResultsThe rtPCR had a sensitivity and specificity of 100% (95% confidence interval = 79–100% and 74–100%, respectively) when testing DNA from other Mycoplasma species or bovine semen spiked with the latter, with a high level of repeatability for within- and between- run replicates. The consistent limit of detection was 0.001 pg/µl M. bovis DNA and between 5.3 × 102 and 7.5 × 102 CFU/ml M. bovis when artificially spiked in semen. DNA extracted using the KingFisher Flex was detected with lower Cq values than the Maxwell 16, but the comparable improvements in sensitivity were mainly associated with non-centrifuged samples (p < 0.001). None of the procedures tested impeded the detection sensitivity of M. bovis in the presence of competitor organisms Acholeplasma laidlawii, Mycoplasma bovigenitalium and Ureaplasma diversum, confirming M. bovis specificity of the polC target.ConclusionsUnder the experimental conditions applied, this rtPCR test efficiently detected M. bovis in extended bovine semen straw samples from DNA extracted using both semi-automated extraction platforms, irrespective of prior centrifugation of extended semen.

An Equine Protein Atlas Highlights Synovial Fluid Proteome Dynamics during Experimentally LPS-Induced Arthritis.

In human proteomics, substantial efforts are ongoing to leverage large collections of mass spectrometry (MS) fragment ion spectra into extensive spectral libraries (SL) as a resource for data independent acquisition (DIA) analysis. Currently, such initiatives in equine research are still missing. Here we present a large-scale equine SL, comprising 6394 canonical proteins and 89,329 unique peptides, based on data dependent acquisition analysis of 75 tissue and body fluid samples from horses. The SL enabled large-scale DIA-MS based quantification of the same samples to generate a quantitative equine protein distribution atlas to infer dominant proteins in different organs and body fluids. Data mining revealed 163 proteins uniquely identified in a specific type of tissue or body fluid, serving as a starting point to determine tissue-specific or tissue-type-specific proteins. We showcase the SL by highlighting proteome dynamics in equine synovial fluid samples during experimental lipopolysaccharide-induced arthritis. A fuzzy c-means cluster analysis pinpointed SERPINB1, ATRN, NGAL, LTF, MMP1, and LBP as putative biomarkers for joint inflammation. This SL provides an extendable resource for future equine studies employing DIA-MS.

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 (&amp;lt;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.

Glucose Oxidase-Based Glucose Biosensing with a Simple Dual Ag/AgCl Probe Conductivity Readout.

We describe a conductometric assay of the enzymatic conversion of glucose to gluconic acid by dissolved glucose oxidase (GOx), using the generation of proton and gluconate from the reaction product dissociation for glucose detection. Simple basics of ionic conductivity, a silver/silver chloride wire pair, and a small applied potential translate glucose-dependent GOx activity into a scalable cell current. Enzyme immobilization and complex sensor design, involving extra nanomaterials or microfabrication of electrode structures, are entirely avoided, in contrast to all modern electrochemical glucose biosensors. Assay calibration showed a response linearity up to 500 μM, with a sensitivity of about 1.3 nA/μM. Selectivity tests excluded signals from sugars other than glucose, and glucose quantifications with recovery rates close to 100% were reached with a model sample and a beverage. Easy use of elementary physicochemical phenomena and a satisfactory performance are assets of the proposed non-amperometric glucose biosensing strategy. Assay integration into a planar dual electrode platform, with or without microfluidic application option, is feasible because of the simplicity of the sensor readout and suggests a route to affordable glucose analysis in beverage, food, and body fluid samples.

The clinical application value of multi-site mNGS detection of patients with sepsis in intensive care units

BackgroundSepsis remains a leading cause of mortality in intensive care units, and rapid and accurate pathogen detection is crucial for effective treatment. This study evaluated the clinical application of multi-site metagenomic next-generation sequencing (mNGS) for the diagnosis of sepsis, comparing its performance against conventional methods.MethodsA retrospective analysis was conducted on 69 patients with sepsis consecutively admitted to the Department of Intensive Care Medicine, Meizhou People’s Hospital. Samples of peripheral blood and infection sites were collected for mNGS and conventional method tests to compare the positive rate of mNGS and traditional pathogen detection methods and the distribution of pathogens. The methods used in this study included a comprehensive analysis of pathogen consistency between peripheral blood and infection site samples. Additionally, the correlation between the pathogens detected and clinical outcomes was investigated.ResultsOf the patients with sepsis, 57.97% experienced dyspnea, and 65.2% had underlying diseases, with hypertension being the most common. mNGS demonstrated a significantly higher pathogen detection rate (88%) compared to the conventional method tests (26%). The pathogen consistency rate was 60% between plasma and bronchoalveolar lavage fluid samples, and that of plasma and local body fluid samples was 63%. The most frequently detected pathogens were gram-negative bacteria, and Klebsiella pneumonia. There were no significant differences in the clinical features between the pathogens.ConclusionmNGS is significantly superior to conventional methods in pathogen detection. There was a notable high pathogen consistency detection between blood and local body fluid samples, supporting the clinical relevance of mNGS. This study highlights the superiority of mNGS in detecting a broad spectrum of pathogens quickly and accurately.Trial registrationNot applicable.

Changing Face of Antimicrobial Resistance: A Cross-Sectional Study

Introduction Infections of sterile body fluids are significant contributors to mortality and morbidity. Accurate species-level identification and understanding of the antimicrobial resistance profiles are crucial for selecting appropriate antimicrobials for empirical and targeted therapy. This study aims to examine the distribution of various bacterial species and their evolved antimicrobial resistance profiles isolated from different body fluids. Methods This retrospective study evaluated 301 body fluid samples collected at a tertiary hospital between January 2023 and December 2023. Samples were gram-stained and cultured on an appropriate media. Bacterial identification and susceptibility testing were performed using the BD Phoenix™ system, with disk diffusion method used for antibiotics not available in the system. Results Microbial growth was detected in 151 cases (50.2%), with a comparable prevalence among males and females (53.0% vs. 46.7%). The mean age was 49.81± 24.1 years. Growth rates were slightly higher in hospital-acquired infections (55.7%) than in community-acquired infections (49.2%). Cerebrospinal fluid showed the highest growth rate at 34.3%. Among the isolates, Streptococcus species were the most common (14.6%), while Escherichia coli was the predominant Gram-negative bacterium (13.2%). Sensitivity was highest with tigecycline (100%) and meropenem (92%), whereas resistance was most notable against ampicillin-sulbactam (100%) and azithromycin (95.2%). Multidrug resistance was identified in 106 isolates (70.2%). Conclusion High antibiotic resistance and multidrug-resistant strains underscore the need for rigorous antibiotic stewardship and improved infection control to address untreatable infections.

A multiplex microbial profiling system for the identification of the source of body fluid and skin samples

Determining the source of body fluids is crucial in forensic investigations, as it provides valuable information about suspects and the nature of the crime. Microbial markers that trace the source of tissues and body fluids based on site specificity and temporal stability are often used effectively for this purpose. In this study, a multiplex system comprising seven microbial markers (Finegoldia magna, Corynebacterium tuberculostearicum, Cutibacterium acnes, Haemophilus parainfluenzae, Streptococcus oralis, Prevotella melaninogenica and Faecalibacterium prausnitzii) was developed to distinguish between skin, saliva, and feces samples. Based on these markers, the system produces electropherograms that are specific for each sample type. We collected 492 samples from six different skin sites (palm, antecubital crease, inguinal crease, cheek, upper back, and toe web space), the buccal mucosa, and stool were collected to further test the system. Beta diversity analysis revealed distinct clustering among the three sample groups. Additionally, skin microenvironment cluster analysis was used to identify skin sites accurately. This analysis classified skin samples into four distinct microenvironments: dry, moist, oily, and foot. Finally, we established a machine learning prediction model based on random forest regression to identify the skin microenvironment, achieving an overall prediction accuracy of 79 %. The multiplex system developed in this study accurately identifies the sources of body fluids, and the skin microenvironment. These findings offer new insights into the application of microbial markers in forensic science.

Development of a multiplex PCR assay and quantification of microbial markers by ddPCR for identification of saliva and vaginal fluid

The identification of biological fluids at crime scenes contributes to crime scene reconstruction and provides investigative leads. Traditional methods for body fluid identification are limited in terms of sensitivity and are mostly presumptive. Emerging methods based on mRNA and DNA methylation require high quality template source. An exploitable characteristic of body fluids is their distinct microbial profiles allowing for the discrimination of body fluids based on microbiome content. Microbial DNA is highly abundant within the body, robust and stable and can persist in the environment long after human DNA has degraded. 16S rRNA sequencing is the gold standard for microbial analysis; however, NGS is costly, and requires intricate workflows and interpretation. Also, species level resolution is not always achievable. Based on the current challenges, the first objective of this study was to develop a multiplex conventional PCR assay to identify vaginal fluid and saliva by targeting species-specific 16S rRNA microbial markers. The second objective was to employ droplet digital PCR (ddPCR) as a novel approach to quantify bacterial species alone and in a mixture of body fluids. Lactobacillus crispatus and Streptococcus salivarius were selected because of high abundance within vaginal fluid and saliva respectively. While Fusobacterium nucleatum and Gardnerella vaginalis, though present in healthy humans, are also frequently found in oral and vaginal infections, respectively. The multiplex PCR assay detected L. crispatus and G. vaginalis in vaginal fluid while F. nucleatum and S. salivarius was detected in saliva. Multiplex PCR detected F. nucleatum, S. salivarius and L. crispatus in mixed body fluid samples while, G. vaginalis was undetected in mixtures containing vaginal fluid. For samples exposed at room temperature for 65 days, L. crispatus and G. vaginalis were detected in vaginal swabs while only S. salivarius was detected in saliva swabs. The limit of detection was 0.06 copies/µl for F. nucleatum (2.5 ×10−9 ng/µl) and S. salivarius (2.5 ×10−6 ng/µl). L. crispatus and G. vaginalis had detection limits of 0.16 copies/µl (2.5 ×10−4 ng/µl) and 0.48 copies/µl (2.5 ×10−7 ng/µl). All 4 bacterial species were detected in mixtures and aged samples by ddPCR. No significant differences were observed in quantity of bacterial markers in saliva and vaginal fluid. The present research reports for the first time the combination of the above four bacterial markers for the detection of saliva and vaginal fluid and highlights the sensitivity of ddPCR for bacterial quantification in pure and mixed body fluids.

Does a postmortem redistribution affect the concentrations of the 7 azaindole-derived synthetic cannabinoid 5F-MDMB-P7AICA in tissues and body fluids following pulmonary administration to pigs?

Many fatal intoxications have been reported in connection with the consumption of newer, highly potent synthetic cannabinoids. Yet, a possible postmortem redistribution (PMR) might complicate reliable interpretation of analytical results. Thus, it is necessary to investigate the PMR-potential of new synthetic cannabinoids. The pig model has already proven to be suitable for this purpose. Hence, the aim of this study was to study the PMR of the synthetic cannabinoid 5F-MDMB-P7AICA and its main metabolite 5F-MDMB-P7AICA-dimethylbutanoic acid (DBA). 5F-MDMB-P7AICA (200 µg/kg body weight) was administered by inhalation to anesthetized and ventilated pigs. At the end of the experiment, the animals were euthanized and stored at room temperature for 3 days. Tissue and body fluid samples were taken daily. Specimens were analyzed after solid phase extraction using a standard addition method and LC–MS/MS, blood was quantified after protein precipitation using a validated method. In perimortem samples, 5F-MDMB-P7AICA was found mainly in adipose tissue, bile fluid, and duodenum contents. Small amounts of 5F-MDMB-P7AICA were found in blood, muscle, brain, liver, and lung. High concentrations of DBA were found primarily in bile fluid, duodenum contents, urine, and kidney/perirenal fat tissue. In the remaining tissues, rather low amounts could be found. In comparison to older synthetic cannabinoids, PMR of 5F-MDMB-P7AICA was less pronounced. Concentrations in blood also appear to remain relatively stable at a low level postmortem. Muscle, kidney, fat, and duodenum content are suitable alternative matrices for the detection of 5F-MDMB-P7AICA and DBA, if blood specimens are not available. In conclusion, concentrations of 5F-MDMB-P7AICA and its main metabolite DBA are not relevantly affected by PMR.

Detection of Beta-lactamase Resistance in Klebsiella pneumoniae Isolated from Different Clinical Sources

186 samples were collected from various pathogenic specimens, including 75 urine samples, 46 cerebrospinal fluid samples, 23 sputum samples, 9 body fluid samples, 14 blood samples, 4 discharge samples, 11 fluid samples from wounds and burns, and 4 bone marrow samples. Accordingly, 23 isolates of K. pneumoniae were identified as follows: 12 from urine samples, three isolates from cerebrospinal fluid, two each from catheters, wounds, and sputum, and one isolate from burns and bronchitis. The 23 isolates under study were tested for their susceptibility to 11 penicillin, cephalosporin, and carbapenem antibiotics, including beta-lactam and other antibiotics. The results showed high resistance of K. pneumoniae to piperacillin at a rate of 91%, and then to ampicillin at a rate of 86.9%, while were sensitive to both imipenem and meropenem, at a rate of 78%. The results showed that 100% of the isolates showed their ability to produce beta-lactamase enzymes as confirmed by the acid and mineral methods. The high percentage of enzymatic production indicates the presence of enzymatic resistance among bacterial isolates under study, which is why the bacteria gave high resistance to antibiotics. The results of the study showed the ability of 14 isolates to produce broad-spectrum beta-lactamase enzymes at a rate of 60%.

A novel approach to cancer screening using the nematode Caenorhabditis elegans-based detection assays.

Early diagnosis of cancer is essential for its effective treatment. Currently, established screening tests are cancer-specific and require screening for each type of cancer separately. The primary objective of cancer research is to develop methods that can detect multiple types of tumors from a single body fluid sample. Multicancer early detection tests aim to detect fragments of circulating tumor DNA, cell-free DNA, circulating microRNAs, or proteins released by cancer cells in the patient's body fluids. However, these tests are not suitable for routine cancer prevention due to their high cost. Therefore, in recent years, cancer screening tests have been developed to detect volatile organic compounds in urine using living organisms, such as nematodes, Caenorhabditis elegans. Measuring only 1 mm in length, C. elegans has the potential to offer a new, efficient, cost-effective, quick, and painless method to detect the presence of tumor. The purpose of this review is to present an overview of the literature on the development and validation of C. elegans-based cancer detection methods. The potential benefits of these assays are significant, as they could become a valuable tool for the early identification and diagnosis of cancer, even though this research is still in its initial stages of development.

MPXV DNA kinetics in bloodstream and other body fluids samples

Since spring 2022, the global epidemiology of the monkeypox virus (MPXV) has changed. The unprecedented increase of human clade II MPXV cases worldwide heightened concerns about this emerging zoonotic disease. We analysed the positivity rates, viral loads, infectiousness, and persistence of MPXV DNA for up to 4 months in several biological samples from 89 MPXV-confirmed cases. Our data showed that viral loads and positivity rates were higher during the first two weeks of symptoms for all sample types. Amongst no-skin-samples, respiratory specimens showed higher MPXV DNA levels and median time until viral clearance, suggesting their usefulness in supporting MPXV diagnosis, investigating asymptomatic patients, and monitoring viral shedding. Infectious virus was cultured from respiratory samples, semen, and stools, with high viral loads and collected within the first 10 days. Notably, only one saliva and one semen were found positive for viral DNA after 71 and 31 days from symptoms, respectively. The focus on bloodstream samples showed the best testing sensitivity in plasma, reporting the overall highest MPXV DNA detection rate and viral loads during the 3-week follow-up as compared to serum and whole-blood. The data here presented can be useful for MPXV diagnostics and a better understanding of the potential alternative routes of its onward transmission.

Analysis of MiR-20b, MIR-197 markers for differentiation between forensic body fluids encountered in sexual assault cases

AbstractIdentifying body fluids can be a critical clue that aids in reconstructing the crime scene. Semen and vaginal fluid identification is crucial, especially in cases of sexual assault. The majority of forensic studies focused on identifying normal body fluids and neglected the expression variation of semen in pathology. To differentiate between vaginal fluids, fertile and infertile semen samples (oligospermia and azoospermia) using miR 20b and miR197. A total of 48 body fluid samples, divided as 16 vaginal fluids, 16 fertile semen, and 16 infertile semen samples (8 with oligospermia and 8 with azoospermia), were collected, and the expression levels of miR-20b and miR-197 were detected by the SYBR Green real-time quantitative PCR technique. Our results showed significant different expression of these miRNAs in normal semen compared to vaginal and infertile semen. Moreover, we designed a model based on Fisher’s discriminant function to forecast the group affiliations of unidentified samples. With three novel equations, we were able to accurately distinguish between semen and vaginal fluid, fertile and infertile semen, and oligospermia and azoospermia semen samples with validation accuracy of 81.3%, 100%, and 100%, respectively. MiR-20b and miR-197 expression levels are efficient and appropriate markers to distinguish semen from vaginal fluid and to differentiate between fertile and infertile semen samples. However, the present study is a preliminary study based on clinical samples, and the potential role of these markers in differentiating real crime scene samples is still unknown, so we recommend further research to investigate these markers expression while using forensic samples.

Defect-rich cypress-leaf-like Cu2+1O@NiF for high-performance non-invasive glucose detection with low detection limit

Non-invasive detection of body fluid glucose concentration is of great significance in the diagnosis and treatment of diabetes, which poses a new challenge to the reduction of the detection limit of glucose sensors. In this work, a novel non-enzymatic glucose sensor (defect-rich cypress-leaf-like Cu2+1O@NiF) was proposed by rapid electrodeposition of cypress-leaf-like Cu2+1O on the surface of nickel foam (NiF) substrate. Due to the high electrochemical active surface area and abundant defects of cypress-leaf-like Cu2+1O providing more catalytic active sites, the prepared non-enzymatic sensor has an ultra-low detection limit (0.9 nM), high sensitivity (88,419.4 μA∙mM−1∙cm−2 (1 nM-0.5 μM) and 2095.5 μA∙mM−1∙cm−2 (1 μM-4.33 mM)) and wide linear range (1 nM-4.33 mM) with good selectivity, reproducibility and stability. Furthermore, the obtained sensors exhibited high detection stability, accuracy and recovery for glucose detection of human body fluid samples, which means that the defect-rich cypress-leaf-like Cu2+1O@NiF electrode is an ideal candidate for non-invasive glucose sensors.

Differentiation Between Abscesses and Unnecessary Intervention Fluid After Pancreas Surgery Using Dual-Energy Computed Tomography.

This study aimed to evaluate the potential of dual-energy computed tomography (CT) to distinguish postoperative ascites, pancreatic fistula, and abscesses. Patients who underwent biliary and pancreatic surgery performed at our institution between June 2021 and February 2022 were included in the study. Postoperative body fluid samples were collected through a drain or percutaneous drainage. These samples were set in a phantom, and imaging data were obtained using dual-energy CT. Image analysis was performed to obtain CT values at each energy in virtual monoenergetic images (VMIs), effective atomic number, iodine map, and virtual non-contrast (VNC) images. VMIs were calculated from 80 and 140 kVp tube data at 10 kV each from 40-140 kV. Additionally, the effective atomic number, iodine map, and VNC images were reconstructed from the material decomposition process using water and iodine as the base material pair. In this study, 25 patients (eight with abscess and 17 with ascites) were included. No significant association was observed between the presence or absence of abscess and malignancy or surgical procedure. The intervention was performed in six of the eight patients with abscesses. In contrast, five of the 17 patients with postoperative ascites required intervention. A significant relationship was observed between the intervention and the presence of an abscess. Significant differences in C-reactive protein values and the incidence of fever were observed between the groups. Only VNC showed a significant difference between the groups. VNC using dual-energy CT could differentiate abscesses from postoperative fluid.

Detection of Free-Living Amoeba in a tertiary care hospital

This study aims to investigate the presence of Free-Living Amoebae (FLA), particularly Acanthamoeba spp., in diverse water systems and body fluids within a hospital. Water and body fluid samples were collected from various locations. Cultivation of FLA was performed using Non-Nutrient Agar (NNA) culture with Escherichia coli, and microscopic observations after staining. Microscopic observations revealed the presence of Acanthamoeba spp. in pre-dialysis tank water and dental flush water samples. The identification of Acanthamoeba in specific water sources underscores the importance of continuous monitoring and preventive strategies to mitigate potential health risks.

Small extracellular vesicles detection using dielectrophoresis-based microfluidic chip for diagnosis of breast cancer

Small extracellular vesicles (sEVs) reflect the genotype and phenotype of original cells and are biomarkers for early diagnosis and treatment monitoring of tumors. Yet, their small size and low density make them difficult to isolate and detect in body fluid samples. This study proposes a novel acDEP-Exo chip filled with transparent micro-beads, which formed a non-uniform electrical field, and finally achieved rapid, sensitive, and tunable sEVs capture and detection. The method requires only 20–50 μL of sample, achieved a limit of detection (LOD) of 161 particles/μL, and can detect biomarkers within 13 min. We applied the chip to analyze the two markers of sEV's EpCAM and MUC1 in clinical plasma samples from breast cancer (BC) patients and healthy volunteers and found that the combined evaluation of sEV's biomarkers has extremely high sensitivity, specificity and accuracy. The present study introduces an alternative approach to sEVs isolation and detection, has a great potential in real-time sEVs-based liquid biopsy.