Early Diagnostic Tool Research Articles

No Clear Clustering Dysbiosis from Salivary Microbiota Analysis by Long Sequencing Reads in Patients Affected by Oral Squamous Cell Carcinoma: A Single Center Study.

Advancements in DNA sequencing technology have facilitated the assessment of the connection between the oral microbiome and various diseases. The aim of the present study was to investigate the salivary microbiota composition employing for the first time in the literature the Oxford Nanopore Technology in patients affected by oral squamous cell carcinoma (OSCC). Unstimulated saliva samples of 31 patients were collected (24 OSCC patients and 7 controls). DNA was extracted using the QIAamp DNA Blood Kit and metagenomic long sequencing reads were performed using the MinION device. In the OSCC group, 13 were males and 11 were females, with a mean age of 65.5 ± 13.9 years; in the control group, 5 were males and 2 were females, with a mean age of 51.4 ± 19.2 years. The border of the tongue was the most affected OSCC site. The microorganisms predominantly detected in OSCC patients were Prevotella, Chlamydia, Tissierellia, Calothrix, Leotiomycetes, Firmicutes and Zetaproteobacteria. This study confirmed the predominance of periodontopathic bacteria in the salivary microbiome in the OSCC group. If a direct correlation between oral dysbiosis and OSCC onset was proven, it could lead to new prevention strategies and early diagnostic tools.

Venous congestion affects neuromuscular changes in pigs in terms of muscle electrical activity and muscle stiffness.

Early detection of venous congestion (VC)-related diseases such as deep vein thrombosis (DVT) is important to prevent irreversible or serious pathological conditions. However, the current way of diagnosing DVT is only possible after recognizing advanced DVT symptoms such as swelling, pain, and tightness in affected extremities, which may be due to the lack of information on neuromechanical changes following VC. Thus, the goal of this study was to investigate acute neuromechanical changes in muscle electrical activity and muscle stiffness when VC was induced. The eight pigs were selected and the change of muscle stiffness from the acceleration and muscle activity in terms of integral electromyography (IEMG) was investigated in three VC stages. Consequently, we discovered a significant increase in the change in muscle stiffness and IEMG from the baseline to the VC stages (p < 0.05). Our results and approach can enable early detection of pathological conditions associated with VC, which can be a basis for further developing early diagnostic tools for detecting VC-related diseases.

Gait Analysis, Metabolic Parameters and Adherence to the Mediterranean Diet in Patients with Type 2 Diabetes Mellitus Compared with Healthy Controls: A Pilot Study.

Patients with type 2 diabetes mellitus (T2DM) are prone to developing diabetic peripheral neuropathy (DPN) with an increased risk of injuries while walking, potentially leading to plantar ulcers. We aimed to assess the early gait changes in T2DM patients without clinical signs of DPN in comparison to age-matched healthy controls (HC). One hundred T2DM patients (78 women, mean age: 66.4 ± 11.5 years) and 50 age-matched HC (34 women, mean age 62.1 ± 7.9 years) were evaluated with the PODOSmart® gait analysis device. Anthropometric and biochemical data, as well as dietary habits were collected for all participants. T2DM patients also completed the Diabetes Distress (DS) self-report validated questionnaire. One patient was excluded from the study due to lack of recent biochemical data. Among the T2DM patients, 88.9% reported little or no DS and 11.1% moderate DS. The T2DM group had higher body mass index, waist circumference, systolic blood pressure, glycated hemoglobin A1c, sodium, white blood cell count, triglycerides and low-density lipoprotein cholesterol, but lower high-density lipoprotein cholesterol than HC (p < 0.05 for all comparisons). The MedDiet score was satisfactory in both groups (p > 0.05). Significant differences were found between the two study groups in gaitline heel off, propulsion speed, foot progression angle, time taligrade phase, stride length, walking speed, angle attack, oscillation speed, pronation-supination toe off and clearance. The T2DM patients without self-reported DS or clinical signs of DPN may exhibit significant differences in several gait parameters analyzed with PODOSmart®. Whether gait analysis can be used as an early diagnostic tool of T2DM complications should be further explored.

CRISPR/Cas-Based Diagnostics in Agricultural Applications.

Pests and disease-causing pathogens frequently impede agricultural production. An early and efficient diagnostic tool is crucial for effective disease management. Clustered regularly interspaced short palindromic repeats (CRISPR) and the CRISPR-associated protein (Cas) have recently been harnessed to develop diagnostic tools. The CRISPR/Cas system, composed of the Cas endonuclease and guide RNA, enables precise identification and cleavage of the target nucleic acids. The inherent sensitivity, high specificity, and rapid assay time of the CRISPR/Cas system make it an effective alternative for diagnosing plant pathogens and identifying genetically modified crops. Furthermore, its potential for multiplexing and suitability for point-of-care testing at the field level provide advantages over traditional diagnostic systems such as RT-PCR, LAMP, and NGS. In this review, we discuss the recent developments in CRISPR/Cas based diagnostics and their implications in various agricultural applications. We have also emphasized the major challenges with possible solutions and provided insights into future perspectives and potential applications of the CRISPR/Cas system in agriculture.

Rapid particle concentration for immunofluorescence enhancement by a surface acoustic wave microchip

At the initial stages of pathogenesis, biomarkers are generated in an insignificant concentration. Hence, the rapid screening for these low-abundance indicators is immensely critical for early disease diagnosis. Enhancement of the immunofluorescence signal emitted from little concentrated biomarkers can aid their detection among other molecules in complex body fluids. Here, we developed an advanced bead-based immunosensing technique combining surface acoustic wave (SAW) for a rapid and sensitive on-chip detection of the diabetic retinopathy biomarker lipocalin 1. As a result, the implementation of the SAW microchip-enabled particle mixing, separation, and concentration in the droplet delivered a 3.6-fold enhanced immunofluorescence signal within 5 s. The integration of fluorescent signal enhancement by SAW, along with the high sensitivity of the immunoassay, featured a limit of detection of 40 pg/mL in 3 min with a sample volume as low as 4 µL. A simple addition of cover glass on the SAW microchip allowed the immuno-sensing process to be completed without any direct sample contact with the interdigital transducer, minimizing the risk of cross-contamination and damage to the chip due to frequent washing. This study demonstrated the potential of the developed SAW device and insights into its utility as an early diagnostic tool.

Evaluation of the performance of InnowaveDX MTB/RIF for the detection of Mycobacterium tuberculosis complex and rifampicin resistance

Objective: To evaluate the performance of Mycobacterium tuberculosis and rifampicin resistance mutation detection kit (InnowaveDX MTB/RIF, referred to as "InnowaveDX") in diagnosing tuberculosis and rifampicin resistance using sputum samples. Methods: From June 19, 2020 to May 16, 2022, patients with suspected tuberculosis were prospectively and consecutively enrolled in Hunan Provincial Tuberculosis Prevention and Control Institute, Henan Provincial Hospital of Infectious Diseases and Wuhan Jinyintan Hospital. A total of 1 328 patients with suspected tuberculosis were finally included. According to the inclusion and exclusion criteria, 1 035 pulmonary tuberculosis patients (357 were confirmed tuberculosis cases and 678 were clinically diagnosed tuberculosis cases) and 180 non-tuberculosis patients were finally included. Sputum samples were collected from all patients for routine sputum smear acid-fastness tests, mycobacterial culture and drug susceptibility testing. Moreover, the diagnostic value of Xpert®MTB/RIF (referred to as "Xpert") and InnowaveDXin detecting tuberculosis and rifampicin resistance was evaluated. Clinical diagnosis and culture results of Mycobacterium tuberculosis were used as reference standards to assess tuberculosis diagnosis, and phenotypic drug sensitivity and Xpert were used as reference standards to assess rifampicin resistance. The sensitivity, specificity, positive predictive value and negative predictive value of the two methods for tuberculosis diagnosis and rifampicin resistance were analyzed. The consistency of the two techniques was analyzed usingkappa test. Results: Taking clinical diagnosis as the reference standard, the detection sensitivity of InnowaveDX [58.0% (600/1 035)] was higher than that of Xpert [51.7% (535/1 035)] in 1035 patients with pulmonary tuberculosis, and the difference was statistically significant (P<0.001). In 270 pulmonary tuberculosis patients with culture-positive pulmonary tuberculosis identified as M.tuberculosis-complex, the positive rates of InnowaveDX and Xpert were both high [99.6%(269/270)and 98.2%(265/270), respectively] and there was no statistical difference. In culture-negative patients with pulmonary tuberculosis, the sensitivity of InnowaveDX was 38.8% (198/511), which was higher than that of Xpert (29.4%, 150/511), and the difference was statistically significant (P<0.001). Taking phenotypic drug-susceptibility testing (DST) as reference, the sensitivity of InnowaveDX to rifampicin resistance was 99.0% (95%CI: 94.7%-100.0%) and the specificity was 94.0%(95%CI: 88.5%-97.4%). With Xpert as the reference, the sensitivity and specificity of InnowaveDX were 97.1% (95%CI: 93.4%-99.1%) and 99.7% (95%CI: 98.4%-100.0%), respectively, and the kappa value was 0.97 (P<0.001). Conclusions: InnowaveDX show a high sensitivity for detecting Mycobacterium tuberculosis, especially in pulmonary tuberculosis patients with a clinical diagnosis and negative culture results. It also showed high sensitivity in detecting rifampicin resistance with DST and Xpert as reference respectively. InnowaveDX is an early and accurate diagnostic tool for TB and drug-resistant TB, particularly suitable for application in low- and middle-income countries.

Role of serum and salivary microRNAs as diagnostic biomarkers in gastric cancer

Introduction: Incidence of Gastric cancer (GC) is increasing alarmingly in the recent past due to changing lifestyle and diet pattern. GC usually has poor prognosis due to delayed diagnosis. Salivary and serum biomarkers are a potential early diagnostic tool for GC. MicroRNAs are promising biomarkers due to their stability in these body fluids and their pivotal role in carcinogenesis. This study helps to determine the role of serum and salivary microRNAs as diagnostic biomarkers in gastric cancer. Materials and Methods: The systematic review was performed as per the Preferred Reporting Items for Systematic reviews and Meta-Analyses (PRISMA) guidelines. The population, intervention, comparison, outcomes, and study (PICOS) framework was used as a strategy for this review. The data were retrieved from online databases such as Medline (PubMed), Web of Science, Scopus, and SpringerLink. Risk of bias and applicability concerns were evaluated using the four domains of QUADAS-2: patient selection, index test, reference standard, procedure and timing. Results: Quality evaluation based on diagnostic accuracy revealed that five studies conducted by Hou et al., Jianhong et al., Kaczor-Urbanowicz et al., Saliminejad et al., and So JBY et al. had the lowest risk of bias, according to a risk of bias chart created using Revman 5.4.1 software. A serum 12-miRNA biomarker assay was validated by the low risk of bias research. Conclusion: This systematic review provides an insight into the possible role and application of an array of miRNAs from various body fluids as a biomarker for the early detection of gastric cancer. The serum 12-miRNA biomarker assay, validated by a low risk of bias research can serve as a potential diagnostic tool to detect gastric cancer. However, the validation of other salivary, serum, and plasma miRNAs as a diagnostic biomarker for gastric cancer mandates further long-term follow-up studies in diverse ethnicity of populations.

Using artificial neural networks for early diagnosis of glaucoma

Purpose: to summarize the experience of the development and application of artificial neural networks (ANW) in early diagnosis of primary open-angle glaucoma (POAG).Material and methods. A total of 690 patients (918 eyes) were tested. The training clinical group consisted of 459 clinical examples (459 eyes), of which 369 eyes had an initial stage of POAG and 90 eyes had no glaucoma. The testing clinical group was represented by 131 examples (131 eyes), of which 110 eyes belonged to patients with POAG and 21 eyes were without glaucoma. The final diagnostic testing using ANW was conducted on 328 eyes with the diagnosis unknown to the researchers, which belonged to people with suspected POAG. The diagnostic complex included an optimally necessary set of research techniques.Results. ANW identified glaucoma in 198 eyes out of those with suspected glaucoma (60.4 %) with 100 % certainty. 76 eyes (23.2 %) were classified as non-glaucoma, or “healthy”; 54 eyes of the suspected glaucoma patients were identified as “doubtful”, whereupon they were retested by a neural network pool consisting of 5 neural networks. According to the results of the retesting, 28 eyes, or 51.9 % of the “doubtful” ones were identified as having glaucoma, whereas 26 eyes (48.1 %) were identified as non-glaucomatous, i. e. healthy.Conclusion. Our experience suggests that artificial neural networks pose no danger to the doctor or the patient and can be viewed as a very convenient tool for early POAG diagnostics.

Assessment of milk and blood enzymes in Holstein cows with subclinical mastitis

The objective of this study was to determine the activities of lactate dehydrogenase (LDH), alkaline phosphatase (ALP), gamma-glutamyl transferase (GGT), aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in the milk and blood of lactating Holstein cows and investigate their association with subclinical mastitis (SCM). This study also aims to evaluate the impact of SCM and its severity based on California mastitis test (CMT) scores on the above mentioned enzymes, as well as to test the efficiency of their use as an early predictive diagnostic tool. Thirty multiparous, lactating dairy Holstein cows were examined using CMT and according to the obtained results classified into four groups; control group 1 (SCM-free = 10, score = 0), group 2 (weak positive = 8, score +1), group 3 (positive = 7, score +2) and group 4 (strong positive = 5, score +3). The milk LDH, ALP, and GGT activities were significantly higher in groups 3 and 4 compared to group 2 and the control group. LDH activity was significantly higher in group 2 compared to the control group. The mean blood LDH and ALP activities in SCM groups were significantly higher than in the control group. ALP blood activity was significantly higher in group 4 compared to group 2. The blood GGT activity was significantly higher in group 4 compared to other groups. There was a significant positive association between milk LDH, ALP, and GGT enzymes as well as the blood enzymes (ALP and GGT) in SCM cows. The results showed that subclinical mastitis causes considerable changes in the activities of enzymes in milk and blood of infected cows.

A Review of Primary. Secondary, and Tertiary Prevention Strategies for Alzheimer’s Disease

This paper explores the non-modifiable and modifiable risk factors for Alzheimer’s, as well as the Alzheimer’s primary prevention, secondary prevention, and tertiary prevention strategies that can be implemented. Alzheimer's is the sixth leading cause of death in the United States and the most expensive chronic disease (Alzheimer’s Association, 2020). It has a detrimental physical, psychological, and financial impact on patients and caregivers (Alzheimer’s Association, 2020). People can reduce their risk of developing Alzheimer’s by eating a healthy diet, using hearing aids for hearing loss, engaging in regular physical activity, spending time with other people, doing mentally stimulating activities, reducing alcohol consumption, and avoiding tobacco (Livingston et al., 2020). Early screening and diagnostic tools are important because they allow people living with Alzheimer’s to establish an effective care plan and to take advantage of treatments, which are most effective in the early stages of the disease (Guzman-Martinez et al., 2021). Some screening and diagnostic tools include biomarkers, scalable screening technologies, and cognitive assessments (Guzman-Martinez et al.,18 2021). Primary and secondary prevention strategies must address the needs of minority populations, who have a higher risk of developing Alzheimer’s compared to the general population (Livingston et al., 2020). Federal public health initiatives, such as the National Alzheimer’s Project Act, the Dementia Prevention Action Plan, and the Behavioral Risk Surveillance System, play an important role in Alzheimer’s public health research, surveillance, and prevention strategies (Chong et al., 2021; Kelley et al., 2018).

Protein Biomarkers in Chronic Kidney Disease in Children-What Do We Know So Far?

Chronic kidney disease (CKD) in children is a major concern of medical care and public health as it is related to high morbidity and mortality due to progression to end-stage kidney disease (ESKD). It is essential to identify patients with a risk of developing CKD to implement therapeutic interventions. Unfortunately, conventional markers of CKD, such as serum creatinine, glomerular filtration rate (GFR) and proteinuria, have many limitations in serving as an early and specific diagnostic tool for this condition. Despite the above, they are still the most frequently utilized as we do not have better. Studies from the last decade identified multiple CKD blood and urine protein biomarkers but mostly assessed the adult population. This article outlines some recent achievements and new perspectives in finding a set of protein biomarkers that might improve our ability to prognose CKD progression in children, monitor the response to treatment, or even become a potential therapeutic target.

Circulating tumour cells for early detection of clinically relevant cancer.

Given that cancer mortality is usually a result of late diagnosis, efforts in the field of early detection are paramount to reducing cancer-related deaths and improving patient outcomes. Increasing evidence indicates that metastasis is an early event in patients with aggressive cancers, often occurring even before primary lesions are clinically detectable. Metastases are usually formed from cancer cells that spread to distant non-malignant tissues via the blood circulation, termed circulating tumour cells (CTCs). CTCs have been detected in patients with earlystage cancers and, owing to their association with metastasis, might indicate the presence of aggressive disease, thus providing a possible means to expedite diagnosis and treatment initiation for such patients while avoiding overdiagnosis and overtreatment of those with slow-growing, indolent tumours. The utility of CTCs as an early diagnostic tool has been investigated, although further improvements in the efficiency of CTC detection are required. In this Perspective, we discuss the clinical significance of early haematogenous dissemination of cancer cells, the potential of CTCs to facilitate early detection of clinically relevant cancers, and the technological advances that might improve CTC capture and, thus, diagnostic performance in this setting.

Power-doppler ultrasonography is an accurate tool for early pregnancy detection in donkeys

Early detection of pregnancy could enhance the reproductive efficiency of donkeys, as the species has a wide range for the first detection of the embryonic vesicle (EV) via B-mode ultrasonography. This study aimed to assess the uterine blood flow of jennies as an early indirect pregnancy diagnostic test. We hypothesized that uterine blood flow is increased, thus, serving as an early pregnancy diagnostic tool. Jennies (n=52) were bred with fresh semen from a fertile jack, and nine jennies were not bred and served as controls; ovulation (Day-0) was confirmed at 12h intervals for all jennies. Power-doppler and B-mode ultrasonography were performed on Days 7 to 10 to assess the uterine horn blood flow and identification of EV, respectively; the latter was repeated on Day 15. Uterine horn blood flow was evaluated using spot meter techniques, measuring mean pixel intensity. Progesterone concentrations were determined with RIA on Days 7 to 10. Data were assessed for normality with Shapiro- Wilk's test and mixed models. Receiving operating curve was used to determine the cut off value for uterine blood flow. Sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) for blood flow on Days 7 to 10 were calculated to identify pregnant jennies and confirmed on Day 15. Significance was set as P≤0.05 and tendency as 0.05<P<0.1. Of 52 bred jennies, 29 (53%) were confirmed pregnant on Day 15. B-modeultrasonography identified 17.2% (5/29) of the EVs on Day 9 and 48.3% (14/29) on Day 10; no EVs were identified on Days 7 to 8. Pregnant jennies (n=29) had greater uterine blood flow in comparison with non-pregnant jennies (144.8±1.5 vs. 139.2±1.3 pixels/unit, n=32) (P=0.007), with an SD of the difference of 6.3 pixels/unit (p=0.008). There was no effect of time (P=0.48) or group x time interaction between (P=0.70) for uterine blood flow. Progesterone concentrations did not change within Days 7 to 10 (P=0.81), and there was no group x time interaction (P=0.49). Progesterone tended to be greater in pregnant than non-pregnant jennies (P=0.07). There was no effect of time (P=0.79), group (P=0.77), or group x time interaction (P=0.92) for uterine horns. Sensitivity from Days-7-to-10 were 86%, 82%, 60%, and 62%, respectively. In contrast, specificity for Days 7 to 10 was 80%, 88%, 78%, and 75%, respectively. The PPV and NPV for Days 7 to 10 were 81% and 85%; 75% and 20%; 35% and 33%; and 30% and 27%; respectively. In conclusion, this study demonstrated the efficacy of power-doppler-ultrasonography as a sensitive and specific tool for early indirect detection of pregnancy in donkeys, whereas standard B-mode first detected a small percentage of pregnancies on Days 9 to 10. Power-doppler could facilitate the breeding management of donkeys, such as timing the induction of luteolysis in non-pregnant jennies at Days 7 to 10 post-ovulation.

A contingency approach for time-cost trade-off in construction projects based on machine learning techniques

PurposeTime-cost trade-off is normal conduct in construction projects when projects are expectedly late for delivery. Existing research on time-cost trade-off strategic management mostly focused on the technical calculation towards the optimal combination of activities to be accelerated, while the managerial aspects are mostly neglected. This paper aims to understand the managerial efforts necessary to prepare construction projects ready for an upcoming trade-off implementation.Design/methodology/approachA preliminary list of critical factors was first identified from the literature and verified by a Delphi survey. Quantitative data was then collected by a questionnaire survey to first shortlist the preliminary factors and quantify the predictive model with different machine learning algorithms, i.e. k-nearest neighbours (kNN), radial basis function (RBF), multiplayer perceptron (MLP), multinomial logistic regression (MLR), naïve Bayes classifier (NBC) and Bayesian belief networks (BBNs).FindingsThe model's independent variable importance ranking revealed that the top challenges faced were the realism of contractual obligation, contractor planning and control and client management and monitoring. Among the tested machine learning algorithms, multilayer perceptron was demonstrated to be the most suitable in this case. This model accuracy reached 96.5% with the training dataset and 95.6% with an independent test dataset and could be used as the contingency approach for time-cost trade-offs.Originality/valueThe identified factor list contributed to the theoretical explanation of the failed implementation in general and practical managerial improvement to better avoid such failure. In addition, the established predictive model provided an ad-hoc early warning and diagnostic tool to better ensure time-cost implementation success.

Role of fT4 and TSH-index fluctuations as early diagnostic tools in milder forms of central hypothyroidism: data from 221 patients with pituitary lesions from a single tertiary center

Searchable abstracts of presentations at key conferences in endocrinology ISSN 1470-3947 (print) | ISSN 1479-6848 (online)

Association of CD4 cell counts and viral load with cystatin C level in children with human immunodeficiency virus (HIV) infection

Background The ease of access to antiretroviral therapy (ART) has improved both survival rate and comorbidities in patients with human immunodeficiency virus (HIV) infection. Impaired kidney function is one of the most common comorbidities of HIV. CD4 and viral load can be used to monitor HIV progression and to determine the effectiveness of ART. The most commonly used estimated-glomerular filtration rate (e-GFR) technique is to use serum creatinine but often causes late detection of kidney dysfunction while serum cystatin increases at the beginning of the GFR decrease. This supports cystatin C serum as an early diagnostic tool to detect kidney function or biomarker early kidney disorders.&#x0D; Objective To evaluate a possible association between serum cystatin C as a marker of kidney function and HIV progression through CD4 levels and viral load.&#x0D; Methods This cross-sectional study was conducted through evaluation of secondary data from medical and laboratory records of pediatric patients who had routine visits to the HIV Clinic at Dr. Hasan Sadikin General Hospital, Bandung, West Java, in January-February 2020.&#x0D; Results Sixty subjects were reviewed in the study. Median cystatin C-based eGFR was 28.1mL/minute/1.73m2. Subjects were categorized by viral load result into &lt;40 and ?40 copies/mL. The median serum cystatin C was significantly higher [3.7 (range 2.61–6.55) mg/L] in the &gt;40 copies/mL viral load group than the &lt;40 copies/mL group [2.4 (range 0.26–13.61) mg/L]. The median absolute CD4 count, CD4 percentage, and cystatin C were 776 (range 7–1644) cells/mm3, 27.5 (range 1.6–57.4) %, and 3 (range 0.26–13.61) mg/L, respectively. There were no significant correlations (r=-0.2; P=0.1) between CD4 and serum cystatin C&#x0D; Conclusion Higher viral load associates with higher cystatin C level, while CD4 shows no correlation to cystatin C. However, patients with low CD4 tend to have increased cystatin C level.&#x0D;

Targeted MRM Quantification of Urinary Proteins in Chronic Kidney Disease Caused by Glomerulopathies.

Glomerulopathies with nephrotic syndrome that are resistant to therapy often progress to end-stage chronic kidney disease (CKD) and require timely and accurate diagnosis. Targeted quantitative urine proteome analysis by mass spectrometry (MS) with multiple-reaction monitoring (MRM) is a promising tool for early CKD diagnostics that could replace the invasive biopsy procedure. However, there are few studies regarding the development of highly multiplexed MRM assays for urine proteome analysis, and the two MRM assays for urine proteomics described so far demonstrate very low consistency. Thus, the further development of targeted urine proteome assays for CKD is actual task. Herein, a BAK270 MRM assay previously validated for blood plasma protein analysis was adapted for urine-targeted proteomics. Because proteinuria associated with renal impairment is usually associated with an increased diversity of plasma proteins being present in urine, the use of this panel was appropriate. Another advantage of the BAK270 MRM assay is that it includes 35 potential CKD markers described previously. Targeted LC-MRM MS analysis was performed for 69 urine samples from 46 CKD patients and 23 healthy controls, revealing 138 proteins that were found in ≥2/3 of the samples from at least one of the groups. The results obtained confirm 31 previously proposed CKD markers. Combination of MRM analysis with machine learning for data processing was performed. As a result, a highly accurate classifier was developed (AUC = 0.99) that enables distinguishing between mild and severe glomerulopathies based on the assessment of only three urine proteins (GPX3, PLMN, and A1AT or SHBG).

Towards in-vivo detection of amyloid−β and tau in human CSF using machine learning based Raman spectroscopy

This paper aims to present initial proof of concept of a non-invasive early diagnostic tool for Alzheimer disease (AD). The approach is based on the identification using Raman spectroscopy and machine learning algorithms of two proteins that are linked with AD and exist in the cerebrospinal fluid (CSF). As demonstrated in previous studies, the concentration of the proteins amyloid-β and tau may indicate the existence of AD. The proteins’ concentration in the CSF signifies the condition of AD. The current study can contribute to the existing body of knowledge by enabling the development of a non-invasive diagnostic tool that may help with early diagnosis of AD.

Genetically engineered bacterial strains constructed as a whole-cell biosensor for specific volatiles identification of infected potato tubers with a soft rot disease

The potato (Solanum tuberosum) is the most globally popular non-cereal crop. During postharvest storage, the potato is susceptible to various diseases. Specifically, the symptoms of tuber soft rot can be identified visibly only after irreversible damage has already been caused. The existing technologies are not optimized for the identification of hidden diseases, thus, an early and sensitive diagnostic tool is needed. An attractive early-stage technology is based on monitoring the volatile organic compounds (VOCs) as indicators of pathogen presence. In this study, genetically engineered bacterial strains were constructed as a whole-cell biosensor for specific volatiles identification of infected potato tubers with soft rot disease. Identified volatiles were chosen (i.e., 1-Octanol, 2-Phenylethyl Alcohol, and 1-Octen-3-ol) as model toxicants and used for the identification of the responsive promoters in the host cell Escherichia coli through RNA sequencing and bioinformatic analysis. Furthermore, the promoter activity was verified by a fusion to luciferase-harboring plasmid. The bioluminescent bacterial cells monitor the changes in the VOC profile, and a signal change is then generated, reflecting the infection status of the potatoes. The efficacy of the recombinant bacteria to monitor potato soft rot disease was investigated in solution, surrounding air near pure volatiles, and around potato tubers contaminated by Pectobacterium. The results demonstrated that the new strains were sensitive to the presence of these VOCs, each to a different extent and in a different environment (solution or surrounding air). All of them except oxyR could detect differences between infected and uninfected potato tubers. The preferred bioreporter to detect Pectobacterium infection in whole potato tubers is dnaK. This strain responds to all tested volatiles in the liquid and gaseous states and has shown the highest response differences as a reaction to the infection presence in the whole potato tubers. Bacterial monitoring of the volatile profile results in identifying specific signatures that will allow quick and accurate detection of the presence of pathogens in stored crops, prior to visible disease symptoms.

Circulating Cell-Free Nucleic Acids as Biomarkers for Diagnosis and Prognosis of Pancreatic Cancer

A lack of reliable early diagnostic tools represents a major challenge in the management of pancreatic cancer (PCa), as the disease is often only identified after it reaches an advanced stage. This highlights the urgent need to identify biomarkers that can be used for the early detection, staging, treatment monitoring, and prognosis of PCa. A novel approach called liquid biopsy has emerged in recent years, which is a less- or non-invasive procedure since it focuses on plasmatic biomarkers such as DNA and RNA. In the blood of patients with cancer, circulating tumor cells (CTCs) and cell-free nucleic acids (cfNAs) have been identified such as DNA, mRNA, and non-coding RNA (miRNA and lncRNA). The presence of these molecules encouraged researchers to investigate their potential as biomarkers. In this article, we focused on circulating cfNAs as plasmatic biomarkers of PCa and analyzed their advantages compared to traditional biopsy methods.