Synthesis and Characterization of a Luminol Based Chemiluminescent Trimeric System.

A luminol based chemiluminescent trimeric system, namely 2,3-dihydro-5,8-di(thiophen-2-yl)phthalazine-1,4-dione (T2B-Lum), bearing thiophene rings as donor units and 2,3-dihydrophthalazine-1,4-dione as an acceptor unit was synthesized in two steps via donor-acceptor-donor approach using two different methods. It was found that T2B-Lum emits chemiluminescent light when exposed to H2O2 in a basic medium, and the presence of substituents and the type of aromatic ring bearing chemiluminescent active group have a direct effect on the compound's sensitivity. Among the members of a large family of metal ions, fluorescent and chemiluminescent T2B-Lum exhibited high sensitivity to Cu2+ and Fe3+ ions. Except for other metal cations (silver(I), cadmium(II), cobalt(II), iron(III), lithium(I), magnesium(II), manganese(II), nickel(II), zinc(II)), it has been observed that T2B-Lum is mostly sensitive to copper(II) ions with a detection limit value of 2.2 × 10- 3 M. On the other hand, T2B-Lum was also found to exhibit a high sensitivity to extremely dilute aqueous solutions (e.g., 1:50.000 dilution) of blood samples, making it a promising candidate for use in forensic applications.

A Quencher-Based Blood-Autofluorescence-Suppression Strategy Enables the Quantification of Trace Analytes in Whole Blood.

Precise quantification of trace components in whole blood via fluorescence is of great significance. However, the applicability of current fluorescent probes in whole blood is largely hindered by the strong blood autofluorescence. Here, we proposed a blood autofluorescence-suppressed sensing strategy to develop an activable fluorescent probe for quantification of trace analyte in whole blood. Based on inner filter effect, by screening fluorophores whose absorption overlapped with the emission of blood, a redshift BODIPY quencher with an absorption wavelength ranging from 600-700 nm was selected for its superior quenching efficiency and high brightness. Two 7-nitrobenzo[c] [1,2,5] oxadiazole ether groups were introduced onto the BODIPY skeleton for quenching its fluorescence and the response of H2 S, a gas signal molecule that can hardly be quantified because of its low concentration in whole blood. Such detection system shows a pretty low background signal and high signal-to-back ratio, the probe thus achieved the accurate quantification of endogenous H2 S in 20-fold dilution of whole blood samples, which is the first attempt of quantifying endogenous H2 S in whole blood. Moreover, this autofluorescence-suppressed sensing strategy could be expanded to other trace analytes detection in whole blood, which may accelerate the application of fluorescent probes in clinical blood test.

Profiling the scent of weathered training aids for blood-detection dogs.

At outdoor crime scenes, cadaver-detection and blood-detection dogs may be tasked with locating blood that is days, weeks or months old. Although it is known that the odour profile of blood will change during this time, it is currently unknown how the profile changes when exposed to the environment. Such variables must be studied in order to understand when the odour profile is no longer detectable by the scent-detection dogs and other crime scene tools should be implemented. In this study, blood was deposited onto concrete and varnished wood surfaces and weathered in an outdoor environment over a three-month period. Headspace samples were collected using solid phase microextraction (SPME) and analysed using comprehensive two-dimensional gas chromatography - time-of-flight mass spectrometry (GC×GC-TOFMS). The chemical odour profiles were compared with the behavioural responses of cadaver-detection and blood-detection dogs during training. Data interpretation using principal component analysis (PCA) and hierarchical cluster analysis (HCA) established that the blood odour could no longer be detected using SPME-GC×GC-TOFMS after two months of weathering on both surfaces. Conversely, the blood-detection dogs had difficulty locating the blood samples after one month of weathering on concrete and after one week of weathering on varnished wood. The scent-detection dogs evaluated herein had not been previously exposed to environmentally weathered blood samples during training. Given that this study was conducted to test the dogs' baseline abilities, it is expected that with repeated exposure, the dogs' capabilities would likely improve. The knowledge gained from this study can assist in providing law enforcement with more accurate training aids for blood-detection dogs and can improve their efficiency when deployed to outdoor crime scenes.

The non-contact detection and identification of blood stained fingerprints using visible wavelength reflectance hyperspectral imaging: Part 1.

Blood is one of the most commonly encountered types of biological evidence found at scenes of violent crime and one of the most commonly observed fingerprint contaminants. Current visualisation methods rely on presumptive tests or chemical enhancement methods. Although these can successfully visualise ridge detail, they are destructive, do not confirm the presence of blood and can have a negative impact on DNA sampling. A novel application of visible wavelength reflectance hyperspectral imaging (HSI) has been used for the detection and positive identification of blood stained fingerprints in a non-contact and non-destructive manner on white ceramic tiles. The identification of blood was based on the unique visible absorption spectrum of haemoglobin between 400 and 500 nm. HSI has been used to successfully visualise ridge detail in blood stained fingerprints to the ninth depletion. Ridge detail was still detectable with diluted blood to 20-fold dilutions. Latent blood stains were detectable to 15,000-fold dilutions. Ridge detail was detectable for fingerprints up to 6 months old. HSI was also able to conclusively distinguish blood stained fingerprints from fingerprints in six paints and eleven other red/brown media with zero false positives.

The non-contact detection and identification of blood stained fingerprints using visible wavelength hyperspectral imaging: Part II effectiveness on a range of substrates.

Biological samples, such as blood, are regularly encountered at violent crime scenes and successful identification is critical for criminal investigations. Blood is one of the most commonly encountered fingerprint contaminants and current identification methods involve presumptive tests or wet chemical enhancement. These are destructive however; can affect subsequent DNA sampling; and do not confirm the presence of blood, meaning they are susceptible to false positives. A novel application of visible wavelength reflectance hyperspectral imaging (HSI) has been used for the non-contact, non-destructive detection and identification of blood stained fingerprints across a range of coloured substrates of varying porosities. The identification of blood was based on the Soret γ band absorption of haemoglobin between 400 nm and 500 nm. Ridge detail was successfully visualised to the third depletion across light coloured substrates and the stain detected to the tenth depletion on both porous and non-porous substrates. A higher resolution setup for blood stained fingerprints on black tiles, detected ridge detail to the third depletion and the stain to the tenth depletion, demonstrating considerable advancements from previous work. Diluted blood stains at 1500 and 1000 fold dilutions for wet and dry stains respectively were also detected on pig skin as a replica for human skin.

A comparison of visible wavelength reflectance hyperspectral imaging and Acid Black 1 for the detection and identification of blood stained fingerprints.

Bloodstains are often encountered at scenes of violent crime and have significant forensic value for criminal investigations. Blood is one of the most commonly encountered types of biological evidence and is the most commonly observed fingerprint contaminant. Presumptive tests are used to test blood stain and blood stained fingerprints are targeted with chemical enhancement methods, such as acid stains, including Acid Black 1, Acid Violet 17 or Acid Yellow 7. Although these techniques successfully visualise ridge detail, they are destructive, do not confirm the presence of blood and can have a negative impact on DNA sampling. A novel application of visible wavelength hyperspectral imaging (HSI) is used for the non-contact, non-destructive detection and identification of blood stained fingerprints on white tiles both before and after wet chemical enhancement using Acid Black 1. The identification was obtained in a non-contact and non-destructive manner, based on the unique visible absorption spectrum of haemoglobin between 400 and 500nm. Results from the exploration of the selectivity of the setup to detect blood against ten other non-blood protein contaminants are also presented. A direct comparison of the effectiveness of HSI with chemical enhancement using Acid Black 1 on white tiles is also shown.

[The usefulness of fecal tests in colorectal cancer screening]. / Utilidad de las pruebas fecales en el cribado del cáncer colorrectal.

Colorectal cancer is a paradigm of neoplasms that are amenable to preventative measures, especially screening. Currently, to carry this out, there are various strategies that have proven effective and efficient. In countries that have organized population-level screening programs, the most common strategy is fecal occult blood testing. In recent years, new methods have appeared that could constitute viable alternatives in the near future, among which the detection of changes in fecal DNA is emphasized. In this article, we review the most relevant papers on colorectal cancer screening presented at the annual meeting of the American Gastroenterological Association held in Chicago in May 2014, with special emphasis on the medium and long-term performance of strategies to detect occult blood in feces and the first results obtained with fecal DNA testing.

Robust blind color deconvolution and blood detection on histological images using Bayesian K-SVD.

Hematoxylin and Eosin (H&E) color variation among histological images from different laboratories can significantly degrade the performance of Computer-Aided Diagnosis systems. The staining procedure is the primary factor responsible for color variation, and consequently, the methods designed to reduce such variations are designed in concordance with this procedure. In particular, Blind Color Deconvolution (BCD) methods aim to identify the true underlying colors in the image and to separate the tissue structure from the color information. Unfortunately, BCD methods often assume that images are stained solely with pure staining colors (e.g., blue and pink for H&E). This assumption does not hold true when common artifacts such as blood are present, requiring an additional color component to represent them. This is a challenge for color standardization algorithms, which are unable to correctly identify the stains in the image, leading to unexpected results. In this work, we propose a Blood-Robust Bayesian K-Singular Value Decomposition model designed to simultaneously detect blood and extract color from histological images while preserving structural details. We evaluate our method using both synthetic and real images, which contain varying amounts of blood pixels.

The impact of accelerant facilitated fire on blood detection and the efficacy of subsequent soot removal methods.

Previous literature has established that recovering heat damaged body fluids is possible, however with little investigation into the effect of accelerants used in initiating arson fires. This study therefore aimed to determine whether presumptive blood detection was affected by heat damage resulting from accelerant facilitated fires. Another objective was to examine various techniques for removing soot, which is a noted barrier to blood detection. The study focused on blood deposited on household flooring materials, one porous and one nonporous surface: carpet and tile respectively. Samples were burned with butane, petrol, and kerosene then presumptively tested using the Kastle Meyer colourimetric blood detection test. Testing was then repeated following soot removal by either wiping, scraping, or using liquid latex. The "strength" of positive detections was evaluated using a scale based on reaction speed and colour intensity. Results demonstrated that accelerants weakened detection strength, although nearly all samples tested positive overall, and the impact of each accelerant on both surface types was largely similar. It was also discovered that soot removal improved the strength of blood detection results in approximately 69% of carpet and 47% of tile samples, with wiping being the superior method on both surface types. Consequently, introducing this investigative step may be critical to maximizing blood evidence recovery in arson casework. These findings indicate the worth in recovering severely burned items, particularly for evidence as crucial as blood.

Enhancing forensic blood detection using hyperspectral imaging and advanced preprocessing techniques.

Bloodstains are pivotal in forensic investigations as they can provide crucial DNA information about individuals involved in a crime. Traditional methods for bloodstain detection, including chemical tests and forensic lights, have limitations such as non-specificity to human blood and susceptibility to false positives. In forensic blood detection, molecular spectroscopy is crucial for identifying the unique spectral fingerprints of blood, which arise from its molecular composition. Hyperspectral imaging (HSI) leverages this principle by capturing a wide spectrum of light for each pixel in an image, allowing for the detailed analysis of various substances. HSI has emerged as a promising alternative, offering non-contact, rapid, and cost-effective detection of bloodstains by analyzing the visible and near-infrared electromagnetic spectrum. This study explores the application of HSI for blood detection, addressing challenges such as spectral mixing, time-related changes in bloodstain spectra, and data complexity. The study introduces a novel framework to optimize HSI data, enhancing the accuracy and efficiency of bloodstain classification, called the Fast Extraction (FE) framework. It includes two stages. The main method in the first one is the Enhancing Transformation Reduction (ETR) method to reduce the dimension and complexity of the HSI. The second stage contains a compatible classification model to enhance feature extraction and classification. Our approach is validated using the HyperBlood datasets and many evaluation methods, demonstrating superior performance compared to state-of-the-art deep learning models. It provides high accuracy (97%-100 %) for all HSIs, overcoming various difficulties and blood collection times.

Evaluation of Forensic Luminol in Detection of Blood Stains in Instruments Following Dental Treatment.

Background Saliva and blood, being biological materials with a high potential for infectious transmission in dental environments, pose significant risks to dental professionals, assistants, and patients alike. Therefore, practitioners must adopt stringent security measures to ensure patient care, considering all parties as potential carriers of microorganisms capable of causing infectious diseases. Currently, various methods of disinfection and sterilization are employed to maintain the aseptic chain effectively. Having reliable methods for detecting substances in liquids, particularly body fluids, is crucial and highly convenient. Luminol, a chemiluminescent agent widely used in forensic science for detecting minute traces of blood that are invisible to the naked eye, presents itself as a valuable tool. Blood, a major bodily fluid often present in instruments following dental procedures, underscores the importance of its detection. Hence, in this study, luminol was utilized to detect blood traces in dental instruments following dental treatment, both before and after sterilization or disinfection. Objective Blood and saliva splashes, together with highly contagious aerosols, are always a part of dental procedures. The objective of the current study is to detect traces of blood stains on face shields, surgical instruments, and endodontic files using luminol before and after sterilization. Materials and methods Sample size calculation was done with G*Power software (Version 3.1.9.4, Düsseldorf, Germany), and a total of 30 instruments were selected for the study. In the present study, a total of 30 items were collected and utilized, including 14 instruments used after implant placement, 12 endodontic files employed after root canal treatment, and four face shields utilized during these procedures. Meanwhile, a freshly prepared luminol solution was applied to these instruments, and they were viewed in a dark environment both before and after sterilization procedures. Luminescence generated by luminol was observed in the instruments, indicative of the presence of blood not visible to the naked eye. Statistical analysis for both groups was done with IBM SPSS Statistics for Windows, Version 16.0 (Released 2007; SPSS Inc., Chicago, IL, USA). Intragroup comparison was done using the Friedman test, and intergroup comparison was done using the Wilcoxon signed-rank test. Results Blood stains and chemiluminescence were visualized in two out of 10 endodontic files (one #15 K-file and #20 K-files) and two out of four face shields. The intragroup comparison was done using the Friedman test, and it was found to be statistically significant (p < 0.05). Intergroup comparison was done using the Wilcoxon signed-rank test and was found to be statistically insignificant (p > 0.05). Conclusion Following sterilization and disinfection, there were no visual blood stains or chemiluminescence. Therefore, luminol was found to be effective in detecting blood stains in endodontic files, surgical instruments, and face shields, as well as in validating the sterilization and disinfection processes. Hence, sterilization in dentistry stands as a critical measure to guarantee patient safety, halt the dissemination of infections, and uphold exemplary clinical care standards.

An extreme learning machine optimized by differential evolution and artificial bee colony for predicting the concentration of whole blood with Fourier Transform Raman spectroscopy.

Raman spectroscopy, with its advantages of non-contact nature, rapid detection, and minimum water interference, is promising for non-invasive blood detection or diagnosis in clinic applications. However, there is a critical issue that how to accurately analyze blood composition by Raman spectroscopy. In this study, we apply extreme learning machine (ELM) algorithm and a multivariate calibration regression model to analyze the results from Raman spectroscopy and determine the component's concentrations in blood samples, including glucose, cholesterol, and triglyceride. Self-adaption differential evolution artificial bee colony (SADEABC) algorithm was further applied to increase the data's accuracy and robustness. The obtained data for coefficient of determination, root mean square error of calibration, root mean square error of prediction, and relative percent deviation, were 0.9822, 0.3993, 0.3827, and 6.6679 for glucose, 0.9786, 0.2104, 0.2088 and 5.9533 for cholesterol, and 0.9921, 0.2744, 0.3433 and 10.5075 for triglyceride, respectively. Results demonstrated that the model based on SADEABC-ELM show much better prediction data than those models based on the ELM and ABC-ELM.

A Soft Sensor for Bleeding Detection in Colonoscopies.

Colonoscopies allow surgeons to detect common diseases i.e. colorectal cancer, ulcers and other ailments. However, there is a risk of bleeding in the lower gastrointestinal (GI) tract while maneuvering endoscopes. This may be due to perforations, hemorrhaging, polyps, diverticuli or post-biopsy complications. Thus, it is essential for the surgeon to be able to detect bleeding at the site and evaluate the severity of blood leakage. This paper presents a soft sensor that can detect the presence of blood at the bleeding site during colonoscopies. The sensor consists of optical waveguides that interface with a microfluidic channel. Blood flow causes absorption and scattering of incident light that can be picked up by the optical sensing apparatus via light transmission through the waveguide. The surgeon can be alerted when bleeding occurs through a graphical user interface. The device is compact and measures only 1 mm thick. This allows the sensor to be circumferentially mounted onto a colonoscope at different locations. The sensor is able to record the presence of blood as an optical loss, rapidly detect the presence of blood in under 100 milliseconds as it enters the microchannel, and differentiate between gastric fluid and blood through changes in measured optical loss.

Assessment of the invisible blood contamination on nurses' gloved hands during vascular access procedures in a hemodialysis unit.

A prospective study was conducted to assess potential invisible blood contamination on nurses' gloved hands during vascular access procedures using the occult blood detection method in a hemodialysis unit. 60.13% (273/454) of samples tested positive for hemoglobin. These results highlighted the importance of hand hygiene and glove change during hemodialysis access care.

Multifunctional self-driven origami paper-based integrated microfluidic chip to detect CRP and PAB in whole blood.

Gastrointestinal fistula, a complication of gastrointestinal cancer surgery, has a high mortality rate. Detection of both C-reactive protein (CRP) and prealbumin (PAB) is advantageous in the auxiliary diagnosis of postoperative complications. However, traditional detection methods are not capable of on-site rapid detection. In an attempt to overcome these challenges, a multifunctional origami-paper-based device (ePADs) was developed to simultaneously detect CRP and PAB in whole blood. After integration, functionalization, and modification, the electrochemical dual-parameter device was capable of separating blood cells and detecting target analytes. The plasma separation performance revealed a sample diffusion time of 75 s for a whole blood sample volume of 73.3 µL. The efficiency of the device in separating blood cells was 99.91%. Electrochemical results showed that the multifunctional device exhibited linearity between 5 pg mL-1 and 1 µg mL-1 for CRP (R2 = 0.990), and between 10 pg mL-1 and 1 µg mL-1 for PAB (R2 = 0.998). The limits of detection for CRP and PAB were 5 and 10 pg mL-1, respectively (S/N = 3). We also successfully evaluated the accuracy of the dual-parameter device with clinical whole blood samples. Based on these results, the multifunctional device can facilitate clinical detection and provide a new platform for domestic point-of-care testing.

A dataset for evaluating blood detection in hyperspectral images.

The sensitivity of imaging spectroscopy to haemoglobin derivatives makes it a promising tool for detecting blood. However, due to complexity and high dimensionality of hyperspectral images, the development of hyperspectral blood detection algorithms is challenging. To facilitate their development, we present a new hyperspectral blood detection dataset. This dataset, published under an open access license, consists of multiple detection scenarios with varying levels of complexity. It allows to test the performance of Machine Learning methods in relation to different acquisition environments, types of background, age of blood and presence of other blood-like substances. We have explored the dataset with blood detection experiments, for which we have used a hyperspectral target detection algorithm based on the well-known Matched Filter detector. Our results and their discussion highlight the challenges of blood detection in hyperspectral data and form a reference for further works.

A comparison between visible wavelength hyperspectral imaging and digital photography for the detection and identification of bloodstained footwear marks.

One of the first challenges that crime scene examiners have is determining if a substance is blood before performing analysis. Conventional methods of detecting blood involve the use of chemicals and different wavelengths of light in tandem with digital photography. However, these methods are destructive or provide false positives. Visible wavelength hyperspectral imaging (HSI) is a noncontact blood detection method that has been proven to provide accurate and reliable results. A novel application of this technique has been used for the detection and positive identification of bloodstained footwear marks, of different dilutions ranging from undiluted to 1:50 with distilled water, and on a range of substrates, and colors. Comparisons between HSI and conventional digital photography were made using a grading scale and analyzed using Mann-Whitney U-tests. The HSI technique was able to detect a statistically significant greater amount of tread detail on white tiles, laminate, carpet, and blue tiles compared with the digital photography technique, which was only superior on black tiles. Critically, the HSI technique was also able to determine that the footwear marks were made in blood. These results show that HSI will be useful in forensic investigations, where it is known that the perpetrator has walked through the victim's blood and left a trail of footwear marks at the crime scene. Even if the perpetrator had time to clean up afterward resulting in diluted stains, HSI would still be able to detect bloodstained footwear marks with a greater amount of detail compared with digital photography.

Modulating the mixed potential for developing biosensors: Direct potentiometric determination of glucose in whole, undiluted blood.

The growing demand for tools to generate chemical information in decentralized settings is creating a vast range of opportunities for potentiometric sensors, since their combination of robustness, simplicity of operation and cost can hardly be rivalled by any other technique. In previous works, we have shown that the mixed potential of a Pt electrode can be controlled with analytical purposes using a coating of Nafion, thus providing a way to develop a potentiometric biosensor for glucose. Unfortunately, the linear range of this device did not match the relevant clinical range for glucose in blood. This work presents a novel strategy to control the mixed potential that allows the development of a potentiometric biosensor for the direct detection of glucose in whole, undiluted blood without any sample pretreatment. By changing the ionomer, the analytical response can be tuned, shifting the linear range while keeping the sensitivity. Aquivion, a polyelectrolyte from the same family as Nafion, is used to stabilize the mixed potential of a platinized paper-based electrode, to entrap the enzyme and to reduce the interference from negatively charged species. Factors affecting the generation of the signal and the principle of detection are discussed. Optimization of the biosensor composition was achieved with particular focus on the characterization of the linear range and sensitivity. The accurate measurement of blood sugar levels in a single drop of whole blood with excellent recovery is presented.

Fecal Hemoglobin Concentration, a Good Predictor of Risk of Advanced Colorectal Neoplasia in Symptomatic and Asymptomatic Patients.

Background: Periodical fecal immunochemical testing (FIT) is a cost-effective strategy in colon cancer screening programmes. FIT is also used as a diagnostic test in symptomatic patients, but data, are scarce. Aim: To determine the association between FIT-Hb concentration and the risk of advanced neoplasia (AN) detected in colonoscopy in two different populations. Methods: The outcomes of colonoscopies performed after a positive FIT (>117 ng/ml) (Sentinel Gold test) result were analyzed in patients included within a population-based CRC screening programme (screening group) and, as diagnostic evaluation in symptomatic patients (symptomatic group). The study was performed between January 1st, 2014 and October 31, 2016. Data are reported as medians with interquartile ranges or frequencies and percentages. Positive predictive value (PPV) at arbitrary fecal hemoglobin concentrations were also reported calculated for AN. Results: We recruited 2742 patients who underwent a colonoscopy procedure, 1515 (53.5%) of them within the CRC screening programme. Patients in the screening group were younger (65.0 ± 3.3 vs. 66.2 ± 13.4 years, p < 0.001) and more frequently male (p < 0.001) vs. the symptomatic group. Colonoscopy found more frequently neoplastic lesions in the screening compared to the symptomatic group (61.9 vs. 44.8% p < 0.001). Hb concentration in FIT was significantly higher in patients with AN compared with patients without AN in both groups (p < 0.001). The age-adjusted risk of AN increased significantly in both groups according to FIT Hb concentration in the Quartile 3 [OR (95% CI): 2.94 (2.33-3.71)] and Quartile 4 [OR: 5.52 (4.36-6.99)]. Males, in both groups showed a higher probability of presenting AN. FIT values were higher for left- than for right-sided AN in the screening, but not in the symptomatic group. Positive predictive values for AN were higher in the screening group in positive FIT tests (range 43.9-70.5%; 117 to >1,000 ng/ml) compared to those in the symptomatic group (36.3-52.5%). Similar trends were observed for cancer diagnosis alone. Conclusions: Male gender, age, and FIT Hb concentration are predictors of risk of advanced adenoma and colorectal cancer and can be used to prioritize colonoscopy in patients with suspected advanced neoplasia, both in screening and in symptomatic patients.

Evaluation of Bluestar® Forensic Magnum and Other Traditional Blood Detection Methods on Bloodstained Wood Subjected to a Variety of Burn Conditions.

Accurate blood detection is a primary concern for forensic scientists, especially in highly compromised situations. In this study, blood was added to wood blocks and subjected to a variety of fire treatments: the absence or presence of accelerant, burn time (1, 3, or 5 min), and extinguishment method (smothering or dousing with water). Burned blocks were given a qualitative burn score, followed by removal of half of the char from each block and subsequent testing of each half for blood using luminol (13% positive; n = 96), Bluestar® Forensic Magnum (5.2% positive; n = 96), and combined phenolphthalein tetramethylbenzidine test (0% positive; n = 192). Luminol and Bluestar® Forensic Magnum performed similarly, both outperforming PTMB. Additionally, positive results were more likely from samples that were smothered, had a low burn score, and had more concentrated blood solutions (neat or 1:2). Overall, it is extremely unlikely that blood would be detected on combustible substrates exposed to direct fire.