Application of FTIR two-dimensional correlation spectroscopy (2D-COS) analysis in characterizing environmental behaviors of microplastics: A systematic review.

Microplastics (MPs) are ubiquitous in the environment, continuously undergo aging processes and release toxic chemical substances. Understanding the environmental behaviors of MPs is critical to accurately evaluate their long-term ecological risk. Generalized two-dimensional correlation spectroscopy (2D-COS) is a powerful tool for MPs studies, which can dig more comprehensive information hiding in the conventional one-dimensional spectra, such as infrared (IR) and Raman spectra. The recent applications of 2D-COS in analyzing the behaviors and fates of MPs in the environment, including their aging processes, and interactions with natural organic matter (NOM) or other chemical substances, were summarized systematically. The main requirements and limitations of current approaches for exploring these processes are discussed, and the corresponding strategies to address these limitations and drawbacks are proposed as well. Finally, new trends of 2D-COS are prospected for analyzing the properties and behaviors of MPs in both natural and artificial environmental processes.

Physicochemical characteristics of airborne microplastics of a typical coastal city in the Yangtze River Delta Region, China.

Airborne microplastics (MPs) are important pollutants that have been present in the environment for many years and are characterized by their universality, persistence, and potential toxicity. This study investigated the effects of terrestrial and marine transport of MPs in the atmosphere of a coastal city and compared the difference between daytime and nighttime. Laser direct infrared imaging (LDIR) and polarized light microscopy were used to characterize the physical and chemical properties of MPs, including number concentration, chemical types, shape, and size. Backward trajectories were used to distinguish the air masses from marine and terrestrial transport. Twenty chemical types were detected by LDIR, with rubber (16.7%) and phenol-formaldehyde resin (PFR; 14.8%) being major components. Three main morphological types of MPs were identified, and fragments (78.1%) are the dominant type. MPs in the atmosphere were concentrated in the small particle size segment (20-50 µm). The concentration of MPs in the air mass from marine transport was 14.7 items/m3 - lower than that from terrestrial transport (32.0 items/m3). The number concentration of airborne MPs was negatively correlated with relative humidity. MPs from terrestrial transport were mainly rubber (20.2%), while those from marine transport were mainly PFR (18%). MPs in the marine transport air mass were more aged and had a lower number concentration than those in the terrestrial transport air mass. The number concentration of airborne MPs is higher during the day than at night. These findings could contribute to the development of targeted control measures and methods to reduce MP pollution.

The identification of potent dual-target monopolar spindle 1 (MPS1) and histone deacetylase 8 (HDAC8) inhibitors through pharmacophore modeling, molecular docking, molecular dynamics simulations, and biological evaluation.

Background: Overexpression of monopolar spindle 1 (MPS1) and histone deacetylase 8 (HDAC8) is associated with the proliferation of liver cancer cells, so simultaneous inhibition of both MPS1 and HDAC8 could offer a promising therapeutic approach for the treatment of liver cancer. Dual-targeted MPS1/HDAC8 inhibitors have not been reported. Methods: A combined approach of pharmacophore modeling and molecular docking was used to identify potent dual-target inhibitors of MPS1 and HDAC8. Enzyme inhibition assays were performed to evaluate the optimal compound with the strongest inhibitory activity against MPS1 and HDAC8. The selectivity of MPH-5 for MPS1 and HDAC8 was assessed on a panel of 68 kinases and other histone deacetylases. Subsequently, molecular dynamics (MD) simulation verified the binding stability of the optimal compound to MPS1 and HDAC8. Ultimately, in vitro cellular assays and in vivo antitumor assays evaluated the antitumor efficacy of the most promising compound for the treatment of hepatocellular carcinoma. Results: Six dual-target compounds (MPHs 1-6) of both MPS1 and HDAC8 were identified from the database using a combined virtual screening protocol. Notably, MPH-5 showed nanomolar inhibitory effect on both MPS1 (IC50 = 4.52 ± 0.21 nM) and HDAC8 (IC50 = 6.07 ± 0.37 nM). MD simulation indicated that MPH-5 stably binds to both MPS1 and HDAC8. Importantly, cellular assays revealed that MPH-5 exhibited significant antiproliferative activity against human liver cancer cells, especially HepG2 cells. Moreover, MPH-5 exhibited low toxicity and high efficacy against tumor cells, and it overcomes drug resistance to some extent. In addition, MPH-5 may exert its antitumor effects by downregulating MPS1-driven phosphorylation of histone H3 and upregulating HDAC8-mediated K62 acetylation of PKM2. Furthermore, MPH-5 showed potent inhibition of HepG2 xenograft tumor growth in mice with no apparent toxicity and presented favorable pharmacokinetics. Conclusion: The study suggests that MPH-5 is a potent, selective, high-efficacy, and low-toxicity antitumor candidate for the treatment of hepatocellular carcinoma.

Implementation of Newborn Screening for Mucopolysaccharidosis Type IVA and Long-Term Monitoring in Taiwan.

BACKGROUND: Mucopolysaccharidosis IVA (MPS IVA) is a rare lysosomal storage disorder arising from a deficiency in N-acetylgalactosamine-6-sulfatase (GALNS). METHODS: From September 2019 to October 2023, a total of 264,843 Taiwanese newborns underwent screening for MPS IVA using dried blood spots and tandem mass spectrometry. RESULTS: Among the 95 referred infants, nine (9%) were confirmed to have MPS IVA (Group 1), 18 (19%) were highly suspected to have MPS IVA (Group 2), 61 (64%) were identified as heterozygotes of MPS IVA (Group 3), and seven (7%) were determined not to have MPS IVA (Group 4). A total of 34 different GALNS (HGNC:4122) gene variants were identified through our MPS IVA newborn screening program. The most prevalent variant was c.857C>T p.(Thr286Met), found in 33 cases (29%), followed by c.953T>G p.(Met318Arg) in 22 cases (19%). Intravenous enzyme replacement therapy (ERT) was initiated in five patients at ages ranging from 0.3 to 1.7 years. The estimated incidence of MPS IVA in this screening program was 3.4 per 100,000 live births. CONCLUSIONS: Due to the progressive nature of MPS IVA, an early diagnosis facilitated by newborn screening and prompt initiation of ERT before irreversible organ damage occurs may result in improved clinical outcomes.

Multifaceted analysis of microplastic pollution dynamics in the Yamuna river: Assessing anthropogenic impacts and ecological consequences.

Microplastics (MPs) are pervasive contaminants that pose significant ecological and human health risks, emerging as one of the most widespread anthropogenic pollutants in natural environments. This study investigates the abundance, characteristics, and distribution of microplastics (MPs) in the Yamuna River, encompassing 29 sampling points across urban, rural, and industrial zones in and around Delhi, Mathura, Haryana, and Agra. Microplastics were identified and quantified using Nile red dye staining and Micro-Raman spectroscopy, with particle size distribution predominantly between 2 µm to 80 µm and the largest detected particle measuring 256.5 µm. The average MPs concentration was 14,717 ± 4444 L-1, with a significant abundance of hazardous polymers such as polyethylene terephthalate (PET), polypropylene (PP), and polystyrene (PS). The study found that MPs were predominantly fragments and films (65.6 %) and fibers (30.6 %), with transparent particles being the most prevalent. The Pollution Load Index (PLI) consistently indicated high-risk levels (PLI > 100) at all sampling sites, highlighting substantial MP contamination. These results underscore the urgent need for continuous monitoring and the development of robust management strategies to address microplastic pollution in the Yamuna River. This study provides valuable insights into MPs spatial distribution and persistence, contributing to an improved understanding of their environmental impacts and guiding future mitigation and regulatory efforts.

Comparison of pristine and aged poly-L-lactic acid and polyethylene terephthalate as microbe carriers in surface water: Displaying apparent differences.

Microplastics (MPs) in water environment are potential carriers for many substances. In this study, pristine degradable poly-L-lactic acid (PLLA) and non-degradable polyethylene terephthalate (PET) MPs and their UV-aged counterparts were exposed to the Yuhangtang River (Y-River). The results showed that the surface morphology and structure of all MPs markedly changed after exposure. Oxygen-containing functional groups and hydrophilicity of aged MPs were higher compared with their pristine counterparts, and further increased after river exposure. The content of extracellular polymers (EPS) of biofilms on MPs increased with the exposure time, and was higher on aged MPs than on pristine ones. Similar results were obtained for most antibiotic resistance genes (ARGs) between pristine and aged MPs, and ARGs were positively related to pathogens. Dominant bacteria on all MPs were Proteobacteria (51.3 %-81.1 %), Chloroflexi (5.2 %-20.9 %) and Firmicutes (0.4 %-15.9 %), which markedly differed from the Y-River community. Aged MPs could enrich more microbes but relatively fewer bacterial species than pristine MPs, and higher enrichment and species diversity were observed on PLLA compared with PET. This study demonstrates that MPs are highly effective carriers for microbes, and the results provide valuable insights for evaluating the potential impact of bio-MPs on aquatic ecological environment.

A microvascularizedin vitroliver model for disease modeling and drug discovery.

Drug discovery for complex liver diseases faces alarming attrition rates. The lack of non-clinical models that recapitulate key aspects of liver (patho)-physiology is likely contributing to the inefficiency of developing effective treatments. Of particular notice is the common omission of an organized microvascular component despite its importance in maintaining liver function and its involvement in the development of several pathologies. Increasing the complexity of in vitro models is usually associated with a lack of scalability and robustness which hinders their implementation in drug development pipelines. Here, we describe a comprehensive liver microphysiological system model comprising stellates, liver-derived endothelial cells and hepatocytes conceived within a scalable and automated platform. We show that endothelial cells self-organize in a microvascular network when co-cultured with stellates in a hydrogel. In a tri-culture, hepatocytes polarize accordingly, with a basolateral side facing blood vessels and an apical side facing bile-canaliculi-like structures. Stellates interact and surround the hollow microvessels. Steatosis was induced by exogenous administration of fatty acids which could be prevented by co-administration of firsocostat. Administration of TGF-ß resulted in an activated stellate cells phenotype which could be prevented by the co-administration of SB-431542. The model was implemented on a microtiter plate format comprising 64 chips which enabled the development of a fully automated, multiplexed fibrosis assay with a robust Z' factor suitable for high-throughput applications.

Exploring the Therapeutic Implications of Co-Targeting the EGFR and Spindle Assembly Checkpoint Pathways in Oral Cancer.

Head and neck cancer (HNC), the sixth most common cancer worldwide, is increasing in incidence, with oral squamous cell carcinoma (OSCC) as the predominant subtype. OSCC mainly affects middle-aged to elderly males, often occurring on the posterior lateral border of the tongue, leading to significant disfigurement and functional impairments, such as swallowing and speech difficulties. Despite advancements in understanding OSCC's genetic and epigenetic variations, survival rates for advanced stages remain low, highlighting the need for new treatment options. Primary treatment includes surgery, often combined with radiotherapy (RT) and chemotherapy (CT). Cetuximab-based chemotherapy, targeting the overexpressed epidermal growth factor receptor (EGFR) in 80-90% of HNCs, is commonly used but correlates with poor prognosis. Additionally, monopolar spindle 1 (MPS1), a spindle assembly checkpoint (SAC) component, is a significant target due to its role in genomic fidelity during mitosis and its overexpression in several cancers. This review explores EGFR and MPS1 as therapeutic targets in HNC, analyzing their molecular mechanisms and the effects of their inhibition on cancer cells. It also highlights the promise of combinatorial approaches, such as microtubule-targeting agents (MTAs) and antimitotic agents, in improving HNC therapies, patient outcomes, and survival rates.

Genotype-phenotype findings in patients with mucopolysaccharidosis II from the Hunter Outcome Survey.

PURPOSE: This study investigated the relationship between mucopolysaccharidosis II (MPS II) iduronate-2-sulfatase gene (IDS) variants and phenotypic characteristics, particularly cognitive impairment, using data from the Hunter Outcome Survey (HOS) registry. METHODS: HOS data for male patients (n = 650) aged ≥5 years at latest cognitive assessment with available genetic data were analyzed. Predefined genotype categories were used to classify IDS variants and report phenotypic characteristics by genotype. RESULTS: At their latest cognitive assessment, 411 (63.2%) of 650 patients had cognitive impairment. Missense variants were the most common MPS II genotype, with about equal frequency for patients with and patients without cognitive impairment. Complete deletions/large rearrangements were associated with cognitive impairment. Cognitive impairment and behavioral issues were most common, and height and weight abnormalities most apparent, in patients with large IDS structural changes. Broadly, missense variants NM-000202.8:c.998C>T p.(Ser333Leu), NM-000202.8:c.1402C>T p.(Arg468Trp), NM-000202.8:c.1403G>A p.(Arg468Gln) and NM-000202.8:c.262C>T p.(Arg88Cys), and splice site variant NM-000202.8:c.257C>T p.(Pro86Leu), were associated with cognitive impairment, and variants NM-000202.8:c.253G>A p.(Ala85Thr), NM-000202.8:c.187 A>G p.(Asn63Asp), NM-000202.8:c.1037C>T p.(Ala346Val), NM-000202.8:c.182C>T p.(Ser61Phe) and NM-000202.8:c.1122C>T were not. CONCLUSION: This analysis contributes toward the understanding of MPS II genotype-phenotype relationships, confirming and expanding on existing findings in a large, geographically diverse population.

Global distribution, drivers, and potential hazards of microplastics in groundwater: A review.

Since microplastics (MPs) were first detected in groundwater, an increasing number of studies have focused on groundwater pollution by MPs. However, knowledge of the global properties of groundwater MPs: distribution, concentration, composition, and morphology remains limited, while potential factors regulating their transport and distribution in groundwater, especially the hydrogeological background and climate warming conditions, have been omitted from most analyses. Furthermore, previous field investigations did not assess the risks posed by groundwater MPs to the environment and to human health, a necessary preliminary to remediation. In this work, to promote future MP pollution studies and remediation policies, we assimilated and synthesized the current knowledge on this topic. We reviewed current data on global groundwater pollution by MPs, analyzed the driving factors of their transport and distribution, and summarized the ecological and health hazards posed by MPs, before discussing current knowledge limits and suggesting perspectives for future work.

Microplastics from polyvinyl chloride agricultural plastic films do not change nitrogenous gas emission but enhance denitrification potential.

The effects of microplastics (MPs) from agricultural plastic films on soil nitrogen transformation, especially denitrification, are still obscure. Here, using a robotized flow-through system, we incubated vegetable upland soil cores for 66 days with MPs from PE mulching film (F-PE) and PVC greenhouse film (F-PVC) and directly quantified the emissions of nitrogenous gases from denitrification under oxic conditions, as well as the denitrification potential under anoxic conditions. The impact of MPs on soil nitrogen transformation was largely determined by the concentration of the additive phthalate esters (PAEs) containing in the MPs. The F-PE MPs with low level of PAEs (about 0.006 %) had no significant effect on soil mineral nitrogen content and nitrogenous gas emissions under oxic conditions. In contrast, the F-PVC MPs with high levels of PAEs (about 11 %) reduced soil nitrate content under oxic conditions, probably owing to promoted microbial assimilation of nitrogen, as the emissions of denitrification products (N2, NO, and N2O) was not affected. However, the F-PVC MPs significantly enhanced the denitrification potential of the soil due to the increased abundance of denitrifiers under anoxic conditions. These findings highlight the disturbance of MPs from agricultural films, particularly the additive PAEs on nitrogen transformation in soil ecosystems.

Deciphering how endogenous mangrove litterfall influences organic matters transformation driven by microbes in sediment with exogenous microplastics inputs.

The effects of endogenous mangrove litterfall (MF) inputs on organic matter transformation in sediment polluted by exogenous microplastics (MPs) were investigated in this work, and their linkage with microbial characteristics was also explored. MF inputs significantly affected organic carbon transformation in MPs-polluted sediment by improving humification, enzymatic activities and carbon utilisation capacity of microbes. Such effects were mainly linked with the enrichment of microbes responsible for organic substance decomposition induced by MF inputs. Indeed, MF addition increased the relative abundance of fermentation- and cellulysis-assoicated bacteria, together with Saprotrophic fungi. Moreover, dissolved matters derived from MF played a non-neglected role in regulating organic carbon transformation in MPs-polluted sediment. Besides, MF addition decreased the complexity of bacterial community network in MPs-polluted sediment but fungal community network became complicated. And the complexity of microbial network was MF amount-dependent. Even though stochastic process was dominated in sediment with or without MF, MF inputs enhanced the relative contribution of determinism and reduced the migration of microbial communities. A strong response of sediment microbes to MF affected sedimentary organic matters transformation driven by microbes. This work uncovered linkages between organic carbon transformation and microbes in sediment with endogenous litterfall and exogenous MPs inputs in mangroves.

Novel biopolymer spray formulation for drug delivery in precision dentistry.

Addressing the growing challenges of periodontal and peri-implant diseases, this study first reports a promising advancement in precision dentistry: an intricately formulated biopolymer spray designed for precise, localized drug delivery during tailored dental procedures. Poly (lactic-co-glycolic acid) (PLGA), recognized for its controlled release, biodegradability, and FDA-approved biocompatibility, forms the core of this formulation. Utilizing the double emulsion method, PLGA microparticles (PLGA-MPs) were loaded with dental antibiotics: sodium amoxicillin (AMX-Na), trihydrate amoxicillin (AMX-Tri), and metronidazole (Met). This antibiotic combination was thoughtfully selected to meet the distinctive requirements of the most impacting dental treatments. The newly developed biopolymer spray underwent thorough in-vitro analysis, revealing an optimized release curve for antibiotics over time, guaranteeing sustained therapeutic efficacy, and dose-dependent efficacy, accommodating personalized treatment approaches. The positive outcomes position the novel biopolymer spray formulation the leaders in advancing localized drug delivery during dental procedures. Moreover, the precise application and the tunable formulation meets the concept of precision medicine: in detail, this formulation represents a significant stride in dental therapeutics, significantly contributing to the predictability of dental implantology.

The presence of microplastics in Baran's newt (Neurergus barani Öz, 1994) and the spotted newt (Neurergus strauchii Steindachner, 1887).

Microplastics (MPs), tiny plastic particles less than 5 mm in size, have emerged as a common and worrying pollutant in marine, freshwater, and terrestrial environments worldwide. In this study, we revealed the microplastic exposure of two endemic newt species for Türkiye. We found that polyethylene terephthalate (PET) was the predominant microplastic polymer type in both species, with the blue fiber shape in particular. We also found that there was a negative correlation between microplastic size and gastrointestinal tract (GIT) weight, but there was no significant difference between body length and GIT weight of both species. Our findings might be surprising as the studied species live in natural spring waters in remote, high-altitude areas. However, the detection of water bottles in their habitats appears to be the reason for their exposure to microplastic pollution. Therefore, reducing the use of single-use plastics is predicted to contribute to the conservation of these endemic newts.

MPS blockade with liposomes controls pharmacokinetics of nanoparticles in a size-dependent manner.

Pharmacokinetics of nanomedicines can be improved by a temporal blockade of mononuclear phagocyte system (MPS) through the interaction with other biocompatible nanoparticles. Liposomes are excellent candidates as blocking agents, but the efficiency of the MPS blockade can greatly depend on the liposome properties. Here, we investigated the dependence of the efficiency of the induced MPS blockadein vitroandin vivoon the size of blocking liposomes in the 100-500 nm range. Saturation of RAW 264.7 macrophage uptake was observed for phosphatidylcholine/cholesterol liposomes larger than 200 nmin vitro. In mice, liposomes of all sizes exhibited a blocking effect on liver macrophages, prolonging the circulation of subsequently administrated magnetic nanoparticles in the bloodstream, reducing their liver uptake, and increasing accumulation in the spleen and lungs. Importantly, these effects became more pronounced with the increase of liposome size. Optimization of the size of the blocking liposomes holds the potential to enhance drug delivery and improve cancer therapy.

Fast formation and maturation enhancement of human liver organoids using a liver-organoid-on-a-chip.

Background: Spatial and functional hepatic zonation, established by the heterogeneous tissue along the portal-central axis of the liver, is important for ensuring optimal liver function. Researchers have attempted to develop reliable hepatic models to mimic the liver microenvironment and analyze liver function using hepatocytes cultured in the developed systems. However, mimicking the liver microenvironment in vitro remains a great challenge owing to the lack of perfusable vascular networks in the model systems and the limitation in maintaining hepatocyte function over time. Methods: In this study, we established a microphysiological system that operated under continuous flush medium flow, thereby allowing the supply of nutrients and oxygen to liver organoids and the removal of waste and release of cytokines therefrom, similar to the function of blood vessels. Results: The application of microphysiological system to organoid culture was advantageous for reducing the differentiation time and enhancing the functional maturity of human liver organoid. Conclusion: Hence, our microphysiological culture system might open the possibility of the miniaturized liver model system into a single device to enable more rational in vitro assays of liver response.

Dual TTK/PLK1 inhibition has potent anticancer activity in TNBC as monotherapy and in combination.

Background: Threonine tyrosine kinase (TTK) and polo-like kinase 1 (PLK1) are common essential kinases that collaborate in activating the spindle assembly checkpoint (SAC) at the kinetochore, ensuring appropriate chromosome alignment and segregation prior to mitotic exit. Targeting of either TTK or PLK1 has been clinically evaluated in cancer patients; however, dual inhibitors have not yet been pursued. Here we present the in vitro and in vivo characterization of a first in class, dual TTK/PLK1 inhibitor (BAL0891). Methods: Mechanism of action studies utilized biochemical kinase and proteomics-based target-engagement assays. Cellular end-point assays included immunoblot- and flow cytometry-based cell cycle analyses and SAC integrity evaluation using immunoprecipitation and immunofluorescence approaches. Anticancer activity was assessed in vitro using cell growth assays and efficacy was evaluated, alone and in combination with paclitaxel and carboplatin, using mouse models of triple negative breast cancer (TNBC). Results: BAL0891 elicits a prolonged effect on TTK, with a transient activity on PLK1. This unique profile potentiates SAC disruption, forcing tumor cells to aberrantly exit mitosis with faster kinetics than observed with a TTK-specific inhibitor. Broad anti-proliferative activity was demonstrated across solid tumor cell lines in vitro. Moreover, intermittent intravenous single-agent BAL0891 treatment of the MDA-MB-231 mouse model of TNBC induced profound tumor regressions associated with prolonged TTK and transient PLK1 in-tumor target occupancy. Furthermore, differential tumor responses across a panel of thirteen TNBC patient-derived xenograft models indicated profound anticancer activity in a subset (~40%). Using a flexible dosing approach, pathologically confirmed cures were observed in combination with paclitaxel, whereas synergy with carboplatin was schedule dependent. Conclusions: Dual TTK/PLK1 inhibition represents a novel approach for the treatment of human cancer, including TNBC patients, with a potential for potent anticancer activity and a favorable therapeutic index. Moreover, combination approaches may provide an avenue to expand responsive patient populations.

New control model for autonomous vehicles using integration of Model Predictive and Stanley based controllers.

In this paper, a robust control method is introduced for autonomous vehicle control in different scenarios. Dual controllers have been used in this method to ensure high performance and low errors during the vehicle's trip. The new control system is called Model Predictive and Stanley based controller (MPS), which is an integration of a model predictive controller and a Stanley controller. Each of these two controllers has its drawbacks and weaknesses. The proposed method tries to overcome these points and come up with a high-performance control system. This hybrid way of combining two of the famous controllers has the benefit of using the best part of each one and trying to enhance the other part. The MPS is tested for both path-following and vehicle control in different scenarios and on both straight and curved roads. This controller has shown high performance and flexibility to deal with different scenarios of autonomous driving. The results are compared to previous types of controllers, and the proposed system outperformed these types.

Comprehensive body fluid identification and contributor assignment by combining targeted sequencing of mRNA and coding region SNPs.

Forensic genetic analyses aim to retrieve as much information as possible from biological trace material recovered from crime scenes. While standard short tandem repeat (STR) profiling is essential to individualize biological traces, its significance is diminished in crime scenarios where the presence of a suspect's DNA is acknowledged by all parties. In such cases, forensic (m)RNA analysis can provide crucial contextualizing information on the source level about a trace's composition, i.e., body fluids/tissues, and has therefore emerged as a powerful tool for modern forensic investigations. However, the question which of several suspects contributed a specific component (body fluid) to a mixed trace cannot be answered by RNA analysis using conventional methods. This individualizing information is stored within the sequence of the mRNA transcripts. Massively parallel sequencing (MPS) represents a promising alternative, offering not only higher multiplex capacity, but also the typing of individual coding region SNPs (cSNPs) to enable the assignment of contributors to mixture components, thereby reducing the risk of association fallacies. Herein, we describe the development of an extensive mRNA/cSNP panel for targeted sequencing on the IonTorrent S5 platform. Our panel comprises 30 markers for the detection of six body fluids/tissues (blood, saliva, semen, skin, vaginal and menstrual secretion), along with 70 linkage-controlled cSNPs for contributor assignment. It exhibited high reliable detection sensitivity with RNA inputs down to 0.75 ng and a conservatively calculated probability of identity of 0.03 - 6 % for individual body fluid-specific cSNP profiles. Limitations and areas for future work include RNA-related allele imbalances, inclusion of markers to correctly identify rectal mucosa and the optimization of specific markers. In summary, our new panel is intended to be a major step forward to interpret biological evidence at sub-source and source level based on cSNP attribution of a body fluid component to a suspect and victim, respectively.