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The identification of biological stains and their tissue resource is an important part of forensic research. Current methods suffer from several limitations including poor sensitivity and specificity, trace samples, and sample destruction. In this study, we profiled the proteomes of menstrual blood, peripheral blood, saliva, semen, and vaginal fluid with mass spectrometry technology. Tissue-enhanced and tissue-specific proteins of each group have been proposed as potential biomarkers. These candidate proteins were further annotated and screened through the combination with the Human Protein Atlas database. Our data not only validates the protein biomarkers reported in previous studies but also identifies novel candidate biomarkers for human body fluids. These candidates lay the foundation for the development of rapid and specific forensic examination methods.

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The scientific literature contains many accounts of application of polymethine dyes, including cyanine dyes, as imaging agents, i.e., "biological stains," for microscopic investigation of biological materials. Currently, many such dyes are used as probes for living cells, i.e., "fluorescent probes." Polymethine dyes are defined here by two criteria. First, they possess a conjugated chain of (2n + 1) sp2-hybridized carbon atoms that connect a terminal π-electron-accepting (π-electron withdrawing) group with a terminal π-electron-donating group. Second, they have an odd number (2n + 3) of π-centers and an even number (2n + 4) of π-electrons in this chain, where n equals the number of -CR2=CR3- groups, usually vinylene groups -CH=CH-. Commercialization of diverse chemical types of many polymethine dyes has been attempted. The dyes that have achieved wide application, however, are limited in number and it is these dyes that are emphasized here. Because these polymethine dyes sometimes have been described by confusing, and sometimes confused, names, we clarify here the chemical categories and names of such dyes for the nonchemist, biomedical end user of such imaging agents. Nevertheless, the nomenclature presented here is not intended to replace the traditional "chromophore" categories of dyestuff chemistry, because the latter are held in place both by wide usage and by venerable authorities, such as the Colour Index.

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OBJECTIVES: To establish a rapid and nondestructive identification method for human body fluid stains and non-biological stains using three-dimensional fluorescence spectroscopy. METHODS: The collected three-dimensional fluorescence spectrum data of human saliva, 3% blood, coffee and Fanta® stains were processed with dimensionality reduction. After wavelet transform, spectral denoising and feature extraction, the classification formula was established. The Fisher discriminant was used for spectrum matching and recognition to establish the analysis method to distinguish stain types. RESULTS: According to the results of data training and comparison, all the recognition accuracies of Fanta®, coffee, saliva and blood were more than 91.39%. Among them, saliva reached 100% recognition accuracy. CONCLUSIONS: Three-dimensional fluorescence spectroscopy is a potential method for rapid and nondestructive identification of biological and non-biological stains.

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INTRODUCTION: Blood and semen stains are the most common biological stains encountered at crime scenes. The washing of biological stains is a common application that perpetrators use to spoil the crime scene. With a structured experiment approach, this study aims to investigate the effects of washing with various chemicals on the ATR-FTIR detection of blood and semen stains on cotton. MATERIALS AND METHODS: On cotton pieces, a total of 78 blood and 78 semen stains were applied, and each group of six stains was immersed or mechanically cleaned in water, 40% methanol, 5% sodium hypochlorite solution, 5% hypochlorous acid solution, 5 g/L soap dissolved pure water, and 5 g/L dishwashing detergent dissolved water. ATR-FTIR spectra gathered from all stains and analyzed with chemometric tools. RESULTS AND DISCUSSION: According to performance parameters of developed models, PLS-DA is a powerful tool for discrimination of washing chemical for both washed blood and semen stains. Results from this study show that FTIR is promising for use in detecting blood and semen stains that have become invisible to the naked eye due to washing of the findings. CONCLUSION: Our approach allows blood and semen to be detected on cotton pieces using FTIR combined with chemometrics, even though it is not visible to the naked eye. Washing chemicals also can be distinguished via FTIR spectra of stains.

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Recent advances in next-generation sequencing technologies (NGS) coupled with machine learning have demonstrated the potential of microbiome-based analyses in applied areas such as clinical diagnostics and forensic sciences. Particularly in forensics, microbial markers in biological stains left at a crime scene can provide valuable information for the reconstruction of crime scene cases, as they contain information on bodily origin, the time since deposition, and donor(s) of the stain. Importantly, microbiome-based analyses provide a complementary or an alternative approach to current methods when these are limited or not feasible. Despite the promising results from recent research, microbiome-based stain analyses are not yet employed in routine casework. In this review, we highlight the two main gaps that need to be addressed before we can successfully integrate microbiome-based analyses in applied areas with a special focus on forensic casework: one is a comprehensive assessment of the method's strengths and limitations, and the other is the establishment of a standard operating procedure. For the latter, we provide a roadmap highlighting key decision steps and offering laboratory and bioinformatic workflow recommendations, while also delineating those aspects that require further testing. Our goal is to ultimately facilitate the streamlining of microbiome-based analyses within the existing forensic framework to provide alternate lines of evidence, thereby improving the quality of investigations.

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OBJECTIVES@#To establish a rapid and nondestructive identification method for human body fluid stains and non-biological stains using three-dimensional fluorescence spectroscopy.@*METHODS@#The collected three-dimensional fluorescence spectrum data of human saliva, 3% blood, coffee and Fanta® stains were processed with dimensionality reduction. After wavelet transform, spectral denoising and feature extraction, the classification formula was established. The Fisher discriminant was used for spectrum matching and recognition to establish the analysis method to distinguish stain types.@*RESULTS@#According to the results of data training and comparison, all the recognition accuracies of Fanta®, coffee, saliva and blood were more than 91.39%. Among them, saliva reached 100% recognition accuracy.@*CONCLUSIONS@#Three-dimensional fluorescence spectroscopy is a potential method for rapid and nondestructive identification of biological and non-biological stains.

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RESUMEN Los residuos líquidos producidos al elaborar tinciones biológicas contienen mezclas de compuestos químicos y microorganismos, que generan un elevado impacto ambiental si no son tratados adecuadamente. Por esta razón, en el presente trabajo se evaluaron a Pleurotus ostreatus, Trametes versicolor, Enterobacter xianfangensis, Pseudomonas azotoformans, Pseudomonas sp., Bacillus subtilis y Pseudomonas fluorescens, para el tratamiento de un residuo líquido que contenía colorantes trifenilmetánicos y azóicos, a escala de laboratorio. Inicialmente, se seleccionaron las cepas con menor efecto antagónico y se determinó su potencial para producir las enzimas Lacasa, Manganeso Peroxidasa y Lignino Peroxidasa, al emplear sustratos inductores y mezclas de colorantes. Para el consorcio fúngico/bacteriano la disminución de las unidades de color y demanda química de oxígeno fueron del 99 % y 70 % a las 96 h. La remoción de estos parámetros se relacionó con la interacción positiva e incremento de las poblaciones de hongos, bacterias y la producción de enzimas ligninolíticas, obteniendo valores a las 96 h de 7.0 y 14.0 unidades logarítmicas para hongos y bacterias, con unas actividades enzimáticas de 75 U/L, 205 U/L y 0.63 U/L para Lacasa, MnP y LiP, respectivamente. Con el presente trabajo se demostró que con el uso consorcios fúngicos/bacterianos se incrementa la remoción de colorantes y se disminuye el tiempo de proceso. Sugiriendo que estos microorganismos podrían ser evaluados en plantas de tratamiento que integren diferentes unidades de tratamiento para optimizar la remoción de contaminantes con baja biodegradabilidad.

ABSTRACT The liquid wastes generated when biological stains are prepared, contain a mixture of chemical compounds and microorganisms, with high environmental impact. For this reason, Pleurotus ostreatus, Trametes versicolor, Enterobacter xianfangensis, Pseudomonas azotoformans, Pseudomonas sp., Bacillus subtilis and Pseudomonas fluorescens, were used to evaluate the treatment of a liquid waste containing triphenylmethane and azo dyes, on a laboratory scale. Initially, the strains with less antagonistic effect among them were selected for their potential to produce enzymes as Laccase, Manganese Peroxidase and Lignin Peroxidase. The enzymatic activity was determined by using inducing substrates and dye mixtures. For fungal/bacterial consortium, the decrease in color, Chemical Oxygen Demand and in Biochemical Oxygen demand was of 99 %, 70 % and 65 % at 96 h, respectively. The removal of these parameters was related to the positive interaction between the populations of fungi, bacteria and the production of ligninolytic enzymes, obtaining values of 7.0 and 14.0 logarithmic units for fungi and total bacteria at 96 h with enzymatic activities of 75 U/ L, 205 U/L and 0.63 U/L for Laccase, MnP and LiP. The present work demonstrates that using of fungal/bacterial consortia, the removal of dyes is increased, and the process time is decreased. Suggesting that these microorganisms could be evaluated in treatment plants that integrate different treatment units to optimize the removal of contaminants with low biodegradability.

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Elderly patients may be different from the average population in regard to the treatment of shoulder disorders. Challenges are the decreased quality of bone, tendons and cartilage, decreased blood perfusion and a generally aged biology. The advantages however are the often more realistic expectations and more cautious use of the extremity, and the limited life expectancy of prosthetic implants is a less pressing issue. Local pathologies such as in the AC-joint or long head of the biceps may also in the aged patient be treated with infiltration or arthroscopic means. If however large rotator cuff tears and osteoarthritis are present, (reverse) total shoulder implants are the treatment of choice due to the high reliability and uncomplicated rehabilitation.

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The traditional microscope, together with the "routine" hematoxylin and eosin (H & E) stain, remains the "gold standard" for diagnosis of cancer and other diseases; remarkably, it and the majority of associated biological stains are more than 150 years old. Immunohistochemistry has added to the repertoire of "stains" available. Because of the need for specific identification and even measurement of "biomarkers," immunohistochemistry has increased the demand for consistency of performance and interpretation of staining results. Rapid advances in the capabilities of digital imaging hardware and software now offer a realistic route to improved reproducibility, accuracy and quantification by utilizing whole slide digital images for diagnosis, education and research. There also are potential efficiencies in work flow and the promise of powerful new analytical methods; however, there also are challenges with respect to validation of the quality and fidelity of digital images, including the standard H & E stain, so that diagnostic performance by pathologists is not compromised when they rely on whole slide images instead of traditional stained tissues on glass slides.

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