Multiplexing Technique Research Articles

Measuring Vaccine Responses in the Multiplex Era.

Vaccine studies in psychoneuroimmunology (PNI) provide insight into biopsychosocial interactions and their role in infectious diseases. Methodologies to measure vaccine responses are therefore of critical importance for PNI researchers. In this chapter, traditional and modern immunoassays for the assessment of vaccine responses are discussed, highlighting how multiplex techniques provide researchers with greater capacity and opportunity for novel research relating to vaccine outcomes.

Comparison of molecular testing methods for diagnosing non-tuberculous mycobacterial infections.

Rapid and accurate identification of non-tuberculous mycobacteria (NTM) is crucial yet challenging, promoting the development of novel molecular techniques such as amplification-based targeted high-throughput sequencing and metagenomic unbiased high-throughput sequencing. We aimed to evaluate the diagnostic value of these molecular techniques for NTM infection. A total of 115 clinical specimens from patients with confirmed NTM infection were subjected to multiplex polymerase chain reaction detection techniques (multi-PCR), metagenomic Next-Generation Sequencing (mNGS), targeted Next-Generation Sequencing (tNGS), and targeted Nanopore sequencing (tNanopore). Positivity rates and species identification were compared among these techniques. The sensitivity of mNGS, tNGS, and multi-PCR in NTM-infection diagnosis was 44.3%, 42.6%, and 36.5%, respectively, while the sensitivity of the three methods in combination increased to 54.8%. The pathogen identification results of mNGS, tNGS and multi-PCR were matched in 80.6% (25/31) samples at the species level, among which 14 samples (45.2%) was completely matched at the subspecies level. The results of tNanopore, tNGS and mNGS at the species level were completely matched in 73.3% (22/30) samples. These molecular assays demonstrated comparable performance in precisely identifying NTM species in clinical specimens, showing their promising potential as efficient and alternative tools for the rapid diagnosis of NTM disease.

Implementasi dan Pengaruh Time Division Multiplexing (TDM) terhadap Kualitas Transmisi Sinyal

Time Division Multiplexing (TDM) is a multiplexing technique that enables the transmission of multiple signals over a single communication channel by allocating separate time slots for each signal stream. This study aims to analyze the working principles of TDM, examples of its implementation, as well as the benefits and drawbacks of TDM in communication networks. The findings show that TDM can enhance data transmission efficiency through flexible bandwidth allocation, minimizing signal interference and thereby improving signal quality. Due to these advantages, TDM is widely used in fiber optic communication systems, such as telephone and internet networks. Additionally, all data processing is conducted within integrated circuits utilizing Plastic Optical Fiber (POF), allowing TDM to be implemented at a low cost.

Kinetic inductance current sensor for visible to near-infrared wavelength transition-edge sensor readout

Single-photon detectors based on the superconducting transition-edge sensor are used in a number of visible to near-infrared applications, particularly for photon-number-resolving measurements in quantum information science. To be practical for large-scale spectroscopic imaging or photonic quantum computing applications, the size of visible to near-infrared transition-edge sensor arrays and their associated readouts must be increased from a few pixels to many thousands. In this manuscript, we introduce the kinetic inductance current sensor, a scalable readout technology that exploits the nonlinear kinetic inductance in a superconducting resonator to make sensitive current measurements. Kinetic inductance current sensors can replace superconducting quantum interference devices for many applications because of their ability to measure fast, high slew-rate signals, their compatibility with standard microwave frequency-division multiplexing techniques, and their relatively simple fabrication. Here, we demonstrate the readout of a visible to near-infrared transition-edge sensor using a kinetic inductance current sensor with 3.7 MHz of bandwidth. We measure a readout noise of 1.4pA/Hz\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$$1.4\\,{{{\\rm{pA}}}}/\\sqrt{{{{\\rm{Hz}}}}}$$\\end{document}, considerably below the detector noise at frequencies of interest, and an energy resolution of (0.137 ± 0.001) eV at 0.8 eV, comparable to resolutions observed with non-multiplexed superconducting quantum interference device readouts.

The contribution of iron metabolism to heart failure: a haematological and biochemical perspective

Abstract Introduction Heart failure (HF) can be influenced by diverse modulators, such as genes involved in iron metabolism - HFE, SLC40A1 and TMPRSS6, and haematological and biochemical parameters - serum iron, ferritin, transferrin saturation, mean corpuscular volume (MCV) and Red Cell Distribution Width (RDW). Objectives To investigate the association of genetic variants in the HFE gene [C282Y (rs1800562) and H63D (rs1799945)], SLC40A1 (rs1439816 and rs2304704) and TMPRSS6 (rs855791) with haematological parameters and biochemical biomarkers of iron metabolism in HF and in its various forms of presentation [preserved Ejection Fraction-HFpEF and non-preserved Ejection Fraction-HFnpEF (middly reduced + reduced)], in each sex. Methods 301 individuals with HF were studied (141 men and 160 women), with a median age of 80 years (min=31 and max=99) and a median body mass index (BMI) of 25.90 kg/m2 (min=16.50 and max=47.30). Multiplex PCR-ARMS technique was used to analyze the HFE gene variants (C282Y and H63D) and Endpoint Genotyping was used for the remaining variants. Statistical analysis was carried out using SPSS software, version 28.0, with a statistical significance level of p<0.05. Results The presence of indicators of iron deficiency anaemia was verified in the haematological and biochemical parameters in our population of HF. Significant differences were found in serum iron values in the presence of the mutant allele for the three genes studied: 1)in the H63D (p=0.006) for females, and more particularly in females with HFnpEF (p=0.020); 2) in the rs1439816 for males (p=0.035) and more particularly in males with HFpEF (p=0.036); and 3) in the rs855791 for males (p=0.018). As for ferritin, significant differences were found in the presence of the mutant allele for the rs2304704 (p=0.001) in females. For transferrin saturation, significant differences were found in the presence of the mutant allele for the three genes studied: 1) in the H63D (p=0.013) for females, more specifically in females with HFpEF (p=0.043); 2) in the rs1439816 in males (p=0.037); and 3) in the rs855791 in females (p=0.034), particularly in females with HFpEF (p=0.023), and in males (p=0.032), particularly in males with HFpEF (p=0.037). Analyzing the MCV, there were statistically significant differences in the presence of the mutant allele for the rs2304704 (p=0.012) in females, particularly in those with HFpEF (p<0.001). Regarding RDW, significant differences were found in the presence of the mutant allele of the C282Y polymorphism in females with HFpEF (p=0.048). Conclusion Ferropenic anaemia highly prevalent in this population with HF. All the genes analyzed contribute to the modulation of the analyzed parameters/biomarkers, however, the SLC40A1 gene showed greater modulating capacity. Genetic modulation was more relevant in HFpEF women, suggesting that this sex is the most susceptible to genetic modulation.

Spatial analysis by current multiplexed imaging technologies for the molecular characterisation of cancer tissues.

Tumours are composed of tumour cells and the surrounding tumour microenvironment (TME), and the molecular characterisation of the various elements of the TME and their interactions is essential for elucidating the mechanisms of tumour progression and developing better therapeutic strategies. Multiplex imaging is a technique that can quantify the expression of multiple protein markers on the same tissue section while maintaining spatial positioning, and this method has been rapidly developed in cancer research in recent years. Many multiplex imaging technologies and spatial analysis methods are emerging, and the elucidation of their principles and features is essential. In this review, we provide an overview of the latest multiplex imaging techniques by type of imaging and staining method and an introduction to image analysis methods, primarily focusing on spatial cellular properties, providing deeper insight into tumour organisation and spatial molecular biology in the TME.

Immunological sub-phenotypes and response to convalescent plasma in COVID-19 induced ARDS: a secondary analysis of the CONFIDENT trial

BackgroundConvalescent plasma (CP) reduced the mortality in COVID-19 induced ARDS (C-ARDS) patients treated in the CONFIDENT trial. As patients are immunologically heterogeneous, we hypothesized that clusters may differ in their treatment responses to CP.MethodsWe measured 20 cytokines, chemokines and cell adhesion markers using a multiplex technique at the time of inclusion in the CONFIDENT trial in patients of centers having accepted to participate in this secondary study. We performed descriptive statistics, unsupervised hierarchical cluster analysis, and examined the association between the clusters and CP effect on day-28 mortality.ResultsOf the 475 patients included in CONFIDENT, 391 (82%) were sampled, and 196/391 (50.1%) had been assigned to CP. We identified four sub-phenotypes representing 89 (22.8%), 178 (45.5%), 38 (9.7%), and 86 (22.0%) patients. The most contributing biomarkers in the principal component analysis were IL-1β, IL-12p70, IL-6, IFN-α, IL-17A, IFN-γ, IL-13, TFN-α, total IgG, and CXCL10. Sub-phenotype-1 displayed a lower immune response, sub-phenotype-2 a higher adaptive response, sub-phenotype-3 the highest innate antiviral, pro and anti-inflammatory response, and adhesion molecule activation, and sub-phenotype-4 a higher pro and anti-inflammatory response, migration protein and adhesion molecule activation. Sub-phenotype-2 and sub-phenotype-4 had higher severity at the time of inclusion. The effect of CP treatment on mortality appeared higher than standard care in each sub-phenotype, without heterogeneity between sub-phenotypes (p = 0.97).ConclusionIn patients with C-ARDS, we identified 4 sub-phenotypes based on their immune response. These sub-phenotypes were associated with different clinical profiles. The response to CP was similar across the 4 sub-phenotypes.Trial registration: Ethics Committee of the University Hospital of Liège CE 2020/239. Clinicaltrials.gov NCT04558476. Registered 2020-09-11, https://www.clinicaltrials.gov/study/NCT04558476.

High Resolving Power Electrospray Ionization Ion Mobility Spectrometer Based on Fourier Deconvolution Multiplexing.

The resolving power of the drift tube ion mobility spectrometry (IMS) is mainly dependent on the drift length, the drift voltage, the pulse width of an ion gate, and the pressure inside the drift tube. Electrospray ionization (ESI)-IMS is a highly sensitive and reliable technique for the detection and analysis of nonvolatile compounds, and high resolving power is necessary to separate structurally similar compounds. To improve the analytical performance of atmospheric pressure ESI-IMS, the Fourier deconvolution (FD) multiplexing technique is investigated as an effective and convenient means to improve the resolving power as well as the signal-to-noise ratio. By reducing the equivalent ion gate opening width to 5 μs using a typical Tyndall-Powell ion shutter, a high resolving power RP up to 170 can be achieved with a drift length of 12 cm and a drift voltage of 10 kV. Rhodamine 6G (R6G), sodium dodecyl sulfate (SDS), methacycline, oxytetracycline, and ractopamine were evaluated using the FD-ESI-IMS, and mixtures with similar ion mobility can be well separated without prolonging the drift length.

Joint PAPR reduction with channel estimation for OFDM-based VLC systems

Abstract This paper presents a new combination of channel estimation (CE) methods and peak-to-average power ratio (PAPR) reduction techniques for orthogonal frequency division multiplexing (OFDM)-based visible light communication (VLC) systems. We first propose iterative clipping and filtering to minimize the PAPR of optical OFDM signals. Then, we introduce a computationally efficient, high-performance channel estimation approach using block-type pilots with modified clipped signals. The proposed method integrates clipping with three distinct CE techniques, resulting in both significant PAPR reduction and enhanced bit error rate (BER) performance. Among the CE methods evaluated, the expectation-maximization (EM) technique provides better BER results compared to other approaches. Simulation results confirm that the combined use of clipping and advanced CE methods leads to notable improvements in PAPR reduction and BER performance for OFDM-based VLC systems.

Signal processing for enhancing railway communication by integrating deep learning and adaptive equalization techniques.

With the increasing amount of data in railway communication system, the conventional wireless high-frequency communication technology cannot meet the requirements of modern communication and needs to be improved. In order to meet the requirements of high-speed signal processing, a high-speed communication signal processing method based on visible light is developed and studied. This method combines the adaptive equalization algorithm with deep learning and is applied to railway communication signal processing. In this research, the wavelength division multiplexing (WDM) and orthogonal frequency division multiplexing (OFDM) techniques are used, and fuzzy C equalization algorithm is used to softly divide the received signals, reduce signal distortion and interference suppression. The experimental results showed that increasing the step size could reduce the equalization effect, while increasing the modulation parameter will increase the bit error rate. Through deep learning to achieve channel equalization, visible light communication could effectively mitigate multi-path transmission and reflection interference, thereby reducing the bit error rate to the level of 0.0001. Under various signal-to-noise ratios, the system using the channel compensation method achieved the lowest bit error rate. This outcome was achieved by implementing hybrid modulation scheme, including Wavelength division multiplexing (WDM) and direct current-biased optical orthogonal frequency division multiplexing (DCO-OFDM) techniques. It has been proved that this method can effectively reduce the channel distortion when the receiver is moving. This study develops a dependable communication system, which enhances signal recovery, reduces interference, and improves the quality and transmission efficiency of railway communication. The system has practical application value in the field of railway communication signal processing.

Performance Evaluation of NovaplexTM Multiplex Real-Time PCR Assay for Detection of Streptococcus agalactiae Serotypes.

Streptococcus agalactiae, or group B streptococcus (GBS), is a Gram-positive pathogen with an extended track record of colonization in the gastrointestinal and genitourinary tracts. GBS can induce disease in individuals across all age demographics, yet it predominantly triggers infections in neonates and the elderly. Identification of the serotype is vital for effective management of the disease as it provides critical information for clinicians on the cause of the disease. In this study, we evaluated the rapid, simple, and easy-to-adopt multiplex real-time PCR technique, NovaplexTM (NovaPCR). A total of 131 clinical isolates of different serotypes were tested using NovaPCR. Observations revealed that 129 isolates showed the same observations as LA and conventional mPCR. NovaPCR accurately identified serotypes IV and V, which were first classified as serotype Ia in the LA test and mPCR, and the difference between the traditional (LA test and mPCR) and NovaPCR methods is only 1.52%. Accurate serotype identification is helpful for monitoring the epidemics and achieving optimal clinical outcomes, and NovaPCR showed a reliable, fast, easy-to-interpret, and cost-efficient performance in GBS serotyping.

Orthogonality of polarization superposition based on polarization holography.

We propose a polarization superposition orthogonal theory based on tensor polarization holography. Based on this theory, the holographic multiplexing capability can be improved measurably. The orthogonality of polarization waves is characterized by the null reconstruction in polarization holography, achieved through the superposition of multiple basic polarization reference waves. This paper analyzes the orthogonality of linear polarization wave superposition and circular polarization wave superposition using the tensor polarization holography theory. Using the polarized holography multiplexing technique, we experimentally verify the orthogonality of polarization wave superposition. Our experimental results align with the theoretical analysis, indicating potential applications in polarization encoding and decoding by this theory, thereby diversifying optical encryption technology Additionally, we demonstrate that polarization superposition orthogonality holds significant promise for optical control technology.

A rapid economical multiplex PCR-RFLP method for molecular detection and genotyping of Giardia duodenalis clinical isolates

Giardia duodenalis is a common cause of diarrheal illness in regions with limited resources. The demand for rapid and cost-effective detection and genotyping methods in large-scale epidemiological studies and clinical diagnostics is imperative. Hence, we developed a multiplex PCR-RFLP technique targeting the tpi gene of G. duodenalis. The assay successfully screened G. duodenalis positive clinical samples (6.33 %; 36/565). It was also able to categorize the isolates into assemblages A (41.66 %; 13/36) and B (58.33 %; 23/36), as well as into subassemblages: AI (13.8 %; 5/36), AII (27.77 %; 10/36), BIII (36.11 %; 15/36) and BIV (22.22 %; 8/36). High diagnostic sensitivity (94.2 %), specificity (100 %) and accuracy (97.1 %) of the PCR assay were obtained, indicating its reliability for diagnosing giardiasis. Notably, the assay demonstrated close concordance with microscopy (κ=0.85) and reference PCR (κ=0.98) results. The optimized method offers a cost-effective and rapid approach for G. duodenalis detection and genotyping, convenient for epidemiological studies and clinical diagnostics.

Analysis of RhC Antigen Weak Expression Combined with Mimicking Autoanti-Ce and Homologous Anti-Jkb Causing Mismatch

To investigate the reasons for the weak expression of RHCE gene in a patient whose mimicking anti-Ce combined with anti-Jkb caused cross-matching non-combination. ABO, Rh, and Kidd blood group antigens were identified by test tube method and capillary centrifugation. Antibody screening and antibody specificity identification were performed using saline, polybrene and antiglobulin in tri-media association with multispectral cells. RHCE gene sequencing and haploid analysis were performed by multiplex PCR technique and RHCE protein modeling was performed using Swiss-Model. The serum of the patient contained anti-Ce mimicking autoantibodies along with anti-Jkb antibodies. c.48G>C, c.150C>T, c.178C>A, c.201A>G, c.203A>G, and c.307C>T mutations were detected in the RHCE triple-molecule sequencing. A 109 bp insertion sequence was found in intron 2, with fragment loss from intron 5-8. The Rh-group genotype was DCe/DCe , and phenotype was CCDee. Genotyping techniques can assist in deducing the molecular mechanisms of some weakly expressed RhC, c, E, and e in patients' sera to aid in the identification of difficult antibodies and thus ensure the safety of patients' blood transfusion.

Molecular Identification of Normal and Adulterated Meat Samples of Local Markets and Restaurants in Kerbala Province

Background: Species specification of the meat is a highly important field of quality control management in meat consumption and industry. Meat forgery with undeclared sources has become a crucial trouble in many countries as well as in Iraq. It means that raw consumed meat or meat derivatives may include different meat genus; so, the components of meat are not regular like the known marker. The diet has sever effect on public health, religious factor, food safety principles, fair-trades and consumers desire. Aim of the study: The current study was aimed to analysis the species of adulterated meat using molecular technique that targets Mitochondrial Cytochrome cyt b gene by Multiplex PCR for detection of meat species and authentication that provide many forensic and judicial applications for several populations throughout the world. Materials and Methods: The following seventy (70) typical meat and meat derivatives were gathered from neighborhood super markets and eateries in the province of Kerbala: 25 cow, 10 buffalo, 25 sheep, and 10 goats. For meat samples, DNA extraction was done. Next, using species-specific primers of the Mitochondrial Cytochrome cyt b gene for cattle, sheep, goats, and buffalo, the collected DNA was amplified using the Multiplex PCR method. Additionally, the presence of Staphylococcus aureus, one of the microorganisms most commonly linked to food illness, was typically examined in all of the meat samples. Results: Multiplex PCR technique resulting in species-specific products of the amplicon sizes for the DNA prepared samples with the lengths 472bp., 124bp., 585bp., and 330bp. for the flesh of cow, buffalo, sheep, and goats, in that order. In addition to this analysis technique indicated that8 (44.4) % of examined cattle meat samples were mixed with buffalo meat and10 (55.6) % of examined sheep meat samples were mixed with goat meat. As well as, most of examined (normal and mixed) meat samples were contaminated with Staphylococcus aureus bacteria. Discussion: It has been demonstrated that the highly conserved Mitochondrial Cytochrome cyt b gene primers, when used in the multiplex PCR methodology, provide a sensitive, straightforward, adaptable, and dependable method for the investigation of meat samples and meat products. Conclusions: The current study's conclusions state that the “mitochondrial cytochrome cyt b gene” is a useful marker for differentiating between normal and adulterated meat. It also states that Staphylococcus aureus was found in 16/52 (30.76%) of the normal meat samples and 12/18 (66.7%) of the adulterated meat samples, with a significant importance of P=0.046.

Primary ciliary dyskinesia: a case report of double DNAH11 mutant alleles

Primary ciliary dyskinesia (PCD), a rare disorder, is genetically varied. Mutations in proteins involved in the structure, function, or assembly of cilia are known to determine situs inversus, male infertility, and chronic destructive airway disease. PCD is inherited by an autosomal recessive pattern of inheritance in most cases. Nonetheless, patterns of autosomal dominant and X-linked inheritance have been mentioned. A history of recurrent upper and lower respiratory tract infections raised clinical suspicion of primary ciliary dyskinesia in a 10-year-old patient. Genetic tests were performed using next-generation sequencing technology (Illumina NextGen) with the multiplex ligation-dependent probe amplification technique for primary ciliopathies and syndromes subject to differential diagnosis. Genetic testing identified two pathogenic variants, not previously associated with a case report in the literature, c.7727A>G (p.Asp2576Gly) and c.8578G>A (p.Gly2860Ser), within the DNAH11 gene, which is associated with autosomal recessive PCD. The result also reported mutations in other genes involved in autosomal recessive PCD (DNAH8, DNAH9 and ZMYND10), which were classified as variants with uncertain clinical significance. Transmission electron microscopy of respiratory cilia and nasal nitric oxide measurement cannot be used to diagnose PCD in patients with DNAH11 mutations because the structure of cilia is normal, and the levels of NO are not constantly low. High-speed video microscopy analysis can be helpful because DNAH11 mutations cause a distinct phenotype of PCD. Nevertheless, the mutation analysis of various PCD-causing genes remains the easiest to conduct and with good results. Genetic research on PCD has identified a number of significant ciliary genes in recent years, offering fresh perspectives on the molecular processes underlying cilia assembly and function. This facilitates the development of new methods for the diagnosis, prevention, and treatment of PCD. However, because it is a highly complex and heterogeneous disease, the field of gene diagnosis and therapy in PCD is still in its infancy.

W-band photonic-aided reconfigurable multichannel vector mm-wave transmission enabled by the digital subcarrier multiplexing technique.

As multimedia service demands rise, radio-over-fiber (RoF) systems necessitate a flexible wireless resource allocation in multi-user scenarios. Optical wavelength division multiplexing (WDM) can meet multi-user application scenarios but has the problems of high hardware cost and complexity. In this Letter, we propose a photonic-aided reconfigurable multichannel vector millimeter-wave (mm-wave) transmission scheme in the RoF system enabled by the digital subcarrier multiplexing (DSCM) technique. The scheme can flexibly adjust the number and frequency spacing of subcarriers as well as the bandwidth and modulation format allocated to each subcarrier, and it can be applied to multi-user scenarios without increasing the cost and complexity of hardware implementation. The off-line experiments verify the system transmission performance with different numbers, bandwidth, and frequency spacing of subcarriers, fully verifying the feasibility of the scheme. Furthermore, the Letter delves into the effects of the subcarrier number, bandwidth, and frequency spacing on overall system performance, highlighting the potential of this scheme for flexible and cost-effective wireless resource allocation in RoF systems.

Optimizing 20 Gbps of ground-to-satellite free-space optical communication in low earth orbit with hybrid wavelength-mode division multiplexing

High-speed free-space optical (FSO) communication has emerged as a promising technology for low earth orbit (LEO) region in the last decade. In this paper, we simulate 20 Gbps simplex ground-to-satellite with various wavelengths and transmitter beam pointing error of 0.1 to 2.5 urad. The C-band wavelength is chosen, and a non-return-zero (NRZ) pulse generator with 16 channels hybrid wavelength-mode division multiplexing (WDM-MDM) technique employing Hermite-Gaussian (HG) modes is used to vary the pointing error. The effect of beam pointing error and receiver aperture diameter was discussed in this paper. A transmitter beam pointing error of 2.5 urad can work appropriately at distances of 1000 km and 1500 km, with a BER value of 1.41 × 10−6 and 3.56 × 10−5, respectively. Based on the receiver aperture diameter of 100 cm, it successfully achieves a BER value of 2.4 × 10−8 at the LEO region and a clear eye diagram.

A Review on Single-Cell analysis of Macrophages in Cancer

Cancer is characterized by uncontrolled cell growth and spread into surrounding tissue. According to the North American Association of Central Cancer Registries (NAACCR) and the National Center for Health Statistics, there were 1,918,030 cancer cases and a total of 609,360 deaths estimated in the US in 2022. Although the direct cause of cancer is not fully understood, it is recognized that the immune system plays a major role in the development and progression of tumors. Investigating tumor-associated macrophages (TAMs) is crucial for understanding the immunosuppressive role that they may play in the tumor microenvironment, which makes them a target for immunotherapies to combat cancer. However, angiogenesis, tumor initiation and invasion promoting signals prevent gaining deeper insight into the intricate molecular mechanisms involved in TAMs and the tumor microenvironment. While researchers in the past have used population samples to analyze cells, recent efforts have incorporated single-cell technologies to study the complex mixture of cells in a tumor, monitor the effects of immunotherapies and more. With the revolutionary advent of the first single-cell RNA sequencing (scRNA-seq), researchers have since been able to gain insight into single-cell data with unprecedented accuracy and develop new immunotherapies through different single-cell technologies, such as polymethylsiloxane (PDMS) and Multiplexed Ion Beam Imaging (MIBI) multiplexing. In this review, we underline the benefits of adopting single-cell technologies to identify distinct cell types in complex environments (i.e., tumors) and highlight the potential application of these technologies along with various multiplexing techniques to immunotherapies.

Abstract B023: Unravelling fibroinflammatory and immune signatures for pancreatic cancer early detection

Abstract Pancreatic cancer is associated with late-stage diagnosis and poor patient prognosis. The disease is characterised by chronic dysregulated immunity and inflammation. Moreover, chronic pancreatitis is linked with an increased risk of pancreatic cancer. Tissue remodelling during disease progression involves a complex network of cells and a crosstalk between cancer cells, stromal components and immune cells, but it remains poorly explored. Thus, to improve the outcome of patients, there is a need for better understanding of biological changes associated with the initiation and progression of pancreatic cancer at the molecular and spatial levels. Using samples obtained from our Accelerated Diagnosis of neuroEndocrine and Pancreatic TumourS (ADEPTS) biobank, we have evaluated blood samples for the discovery of non- invasive biomarkers to discriminate patients with pancreatic cancer from symptomatic patients without cancer. In this pilot study, the transcriptome of peripheral blood mononuclear cells (PBMCs), isolated using a ficoll gradient was explored to identify PDAC-associated mRNA signatures. Moreover, the Olink® Explore Inflammation and Oncology panels were performed in a cohort of 40 patients with early stage (I-II) PDAC, 40 chronic pancreatitis samples and 40 control patients with benign symptomatic upper GI diseases. To validate and enrich these results, a panel of inflammatory and immune markers including immunoglobulins, cytokines and chemokines was also run using multiplex techniques. Symptoms observed in our cohort were also analysed together with clinical (e.g diabetic status, biochemistry profile) and demographic data (age, sex, ethnicity, post code etc). We have identified mRNA transcriptome signatures and differentially expressed proteins associated with inflammatory and immunological processes in PDAC compared to benign symptomatic samples. Our results offer potential diagnostic utility for early-stage disease. Additionally, our unique cohort of patient samples comprising of those with confirmed PDAC and those presenting to clinic with benign disease and similar non-specific symptoms, allows for the development of non-invasive diagnostic tests with better translational applicability. Future work will involve the validation of these findings in a larger patient cohort and the use of NanoString GeoMx® Digital Spatial Profiler to perform whole transcriptome and proteome profiling in human chronic pancreatic and pancreatic cancer tissue samples. Citation Format: Kyra Fraser, Maria Rosado, Stephen P Pereira, Pilar Acedo. Unravelling fibroinflammatory and immune signatures for pancreatic cancer early detection [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Advances in Pancreatic Cancer Research; 2024 Sep 15-18; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(17 Suppl_2):Abstract nr B023.