Ml Of Serum Research Articles

Abstract 6596: Genomic and proteome analysis using preoperative cell free DNA and extracellular vesicles of lung cancer patients

Abstract Background: The detection of circulating cell free DNA (cfDNA) has been examined as a predictor of postoperative recurrence and survival in lung cancer patients. Extracellular vesicles (EVs) have potent for searching a new target to cancers. Objective: To analyze the association between cfDNA/EVs and prognosis after lung cancer surgery. Methods: We retrospectively examined patient background, disease-free survival, overall survival, and preoperative cfDNA in lung cancer patients who underwent surgery at Kanazawa University Hospital between January 2017 and May 2020, in whom preoperative chemotherapy was not administered. We extracted cfDNA from 4 ml plasma within 7 days before surgery and measured its concentration. EVs were extracted from 500 μl serum via Tim4 protein. Proteins included in EVs were analyzed by mass spectrometry. Results: 285 patients were included, 179 males and 106 females, median age 71(36-88) years, and 8 patients had SCLC. EGFRm- positive lung cancer was included in 93 cases (32.6%). The pStage0/l/ll/lll/lV of NSCLC was 7/187/42/36/6 respectively, and the pStagel/ll/lll of SCLC was 4/1/3 respectively. The mDFS of NSCLC was 14.6 (7.8-29.7)/21.5 (8.5-NA) months in pStage lll/lV, respectively. mOS was 53.3 (28.7-NA) months in pStage lll and was not achieved in other stages. The mDFS of SCLC was 19.8 (1.9-NA)/NA/5.7 (1.9-NA) months in pStage l/ll/lll, respectively. mOS was NA/NA/9.3 (9.3-NA) months, respectively. cfDNA was detected in 283 (99.2%) patients, with a concentration of 4.7 (0-60.0) ng/μl. The correlation coefficient between cfDNA concentration and DFS/OS in NSCLC was -0.023 (P=0.70)/-0.014 (P=0.81). The correlation coefficient between cfDNA concentration and DFS/OS for SCLC was -0.86 (P=0.024)/-0.89 (P=0.012). No recurrence was observed in patients in whom cfDNA was not detected.EVs were detectable from pre-operative serum samples, and the proteins included in these were different, whether disease was recurrent or not. Conclusion: The cfDNA concentration did not correlate with DFS/OS in post-operative NSCLC, whereas it correlated negatively with DFS/OS in SCLC. In the presentation, we will further discuss about the concrete genetic alterations and protein expression, based on the cfDNA sequencing and proteomics analysis. Citation Format: Hayato Koba, Hideharu Kimura, Seiji Yano. Genomic and proteome analysis using preoperative cell free DNA and extracellular vesicles of lung cancer patients [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 6596.

A novel Toxoplasma gondii TGGT1_316290 mRNA-LNP vaccine elicits protective immune response against toxoplasmosis in mice.

Toxoplasma gondii (T. gondii) can infect almost all warm-blooded animals and is a major threat to global public health. Currently, there is no effective drug or vaccine for T. gondii. In this study, bioinformatics analysis on B and T cell epitopes revealed that TGGT1_316290 (TG290) had superior effects compared with the surface antigen 1 (SAG1). TG290 mRNA-LNP was constructed through the Lipid Nanoparticle (LNP) technology and intramuscularly injected into the BALB/c mice, and its immunogenicity and efficacy were explored. Analysis of antibodies, cytokines (IFN-γ, IL-12, IL-4, and IL-10), lymphocytes proliferation, cytotoxic T lymphocyte activity, dendritic cell (DC) maturation, as well as CD4+ and CD8+ T lymphocytes revealed that TG290 mRNA-LNP induced humoral and cellular immune responses in vaccinated mice. Furthermore, T-Box 21 (T-bet), nuclear factor kappa B (NF-kB) p65, and interferon regulatory factor 8 (IRF8) subunit were over-expressed in the TG290 mRNA-LNP-immunized group. The survival time of mice injected with TG290 mRNA-LNP was significantly longer (18.7 ± 3 days) compared with the survival of mice of the control groups (p < 0.0001). In addition, adoptive immunization using 300 μl serum and lymphocytes (5*107) of mice immunized with TG290 mRNA-LNP significantly prolonged the survival time of these mice. This study demonstrates that TG290 mRNA-LNP induces specific immune response against T. gondii and may be a potential toxoplasmosis vaccine candidate for this infection.

Shrink polymer based electrochemical sensor for point-of-care detection of prostate-specific antigen

There is a growing but unmet need for point-of-care detection of prostate-specific antigen (PSA) in body fluid which may facilitate early diagnosis and therapy of prostate cancer in a cost-effective and user-friendly way. Low sensitivity and narrow detection range limits applications of point-of-care testing in practice. Here, an immunosensor is first presented based on shrink polymer and integrated into a miniaturized electrochemical platform for detecting PSA in clinical samples. The sensing electrode was prepared by sputtering a gold film on shrink polymer, followed by heating to shrink the electrode to a small size with wrinkles from nano-scale to micro-scale. These wrinkles can be directly regulated by the thickness of the gold film with high specific areas for enhancement of antigen-antibody binding (3.9 times). A distinct difference between electrochemical active surface area (EASA) and response to PSA of shrink electrodes was observed and discussed. The electrode was treated with air plasma and modified with self-assembled graphene to further enhance the sensor's sensitivity (10.4 times). The shrink sensor with gold 200 nm thick integrated into the portable system was validated by a label-free immunoassay for detection of PSA in 20 μL serum within 35 mins. It exhibited a limit of detection of 0.38 fg/mL, the lowest among label-free PSA sensors, and a wide linear response from 10 fg/mL to 1000 ng/mL. Moreover, the sensor demonstrated reliable assay results in clinical serums, comparable to the commercial chemiluminescence instrument, confirming its feasibility for clinical diagnosis.

A microfluidic platform for detection and quantification of two biomarkers for rheumatoid arthritis

Early detection of rheumatoid factors (RF) and cyclic citrullinated peptide antibodies (anti-CCP) has been proven to be effective in diagnosing, and even preventing progression to more advanced-stages, of rheumatoid arthritis (RA), thereby decreasing irreversible damage to joints. Herein a novel, dual-target detection approach was integrated in a 45 mm × 62 mm microfluidic chip that featured a symmetrical design in which magnetic beads surface-coated with fragment-crystallizable regions of immunoglobulin G and biotinylated CCP enabled detection of these two RA biomarkers within 55 min with only 5 μL of serum. This device could therefore be useful for rapid clinical diagnosis of this all-too-common ailment.

Investigating serum extracellular vesicles in Cystic Fibrosis

BackgroundExtracellular vesicles (EVs) are emerging as biomarkers of disease with diagnostic potential in CF. With the advent of highly effective modulator therapy, sputum production is less common and there is a need to identify novel markers of CF disease progression, exacerbation and response to therapies in accessible fluids such as serum. MethodsWe used size exclusion chromatography (SEC) to isolate and characterise EVs from the blood of PWCF of different ages and compared to ultracentrifugation (UC). We used nanoparticle tracking analysis to measure the number of EVs present in serum obtained from children and adults with CF. Mass spectrometry based proteomics was used to characterise protein expression changes between the groups. ResultsEVs were successfully isolated in SEC fractions from 250 µl serum from PWCF in greater numbers (p <0.01) than density ultracentrifugation. There was not a significant difference in EV numbers between young children with CF and controls. However, there was significantly more EVs in adults compared to children (<6yrs) (p < 0.05). EVs from PWCF before and after Kaftrio treatment were also analysed. Significant protein expression changes were observed within all 3 group. The largest changes detected were between children and adults with CF (57 proteins had a 1.5 fold change in expression with 19 significant changes p < 0.05) and PWCF taking Kaftrio (24 significant changes in EV protein expression was observed 12 months post treatment). ConclusionIn this pilot study, we performed an initial characterisation of EVs in serum from PWCF demonstrating the potential of serum EVs for further diagnostic investigation.

Dual Ion-Selective Membrane Deposited Ion-Sensitive Field-Effect Transistor Integrating a Whole Blood Processing Microchamber for In Situ Blood Ion Testing.

Blood ion testing is one of the methods that is commonly used for monitoring the immune status and providing physiological information for disease diagnosis. However, traditional blood ion sensing methods often require well-trained operators to process the whole blood sample and perform the measurements using bulky instruments, making real-time and continuous blood ion sensing at the bedside difficult. To address the above issues, we proposed a dual ion-selective membrane deposited ion-sensitive field-effect transistor (DISM-ISFET) sensor integrating a microchamber to enable on-chip serum extraction and in situ Na+/K+ ion sensing. As a proof of concept, we sequentially dispensed NaCl and KCl solutions at various concentrations on the DISM-ISFET to find out the highest ion sensitivity and selectivity. Next, we also confirm the high red blood cell sedimentation and serum purity using a microchamber. Finally, we evaluated the system performance using nine clinical whole blood samples and compared their Na+/K+ ion-sensing results with a commercial pocket ion meter. To sum up, our results showed that a DISM-ISFET system can successfully extract 200 μL serum from 500 μL whole blood sample and simultaneously achieve Na+/K+ ion sensing. All the sample processes and measurements can be finished within 10 min in a single chip. We envision this compact and easy-to-use system can be potentially used for various medical environments requiring real-time and continuous blood ion monitoring, such as in a hemodialysis room, operation room, and intensive care unit.

Efficacy and Safety of Autologous Serum Therapy in Chronic Spontaneous Urticaria in the Pediatric Population: A Prospective Pilot Study.

Chronic spontaneous urticaria (CSU) in children is mostly spontaneous in onset (57%). Treatment comprises long-term antihistaminic therapy without need for elaborate investigations. A subset of such patients don't respond to conventional treatment and novel therapies to help reduce pill burden is the need of the hour. To determine the efficacy and safety of autologous serum therapy (AST) in pediatric patients with chronic spontaneous urticaria. All pediatric patients, aged between 6-16 years, attended to our OPD from March 2019 to March 2020 were recruited. Clinico-demographic data and baseline investigations of all patients were performed. Two-weekly AST therapy was given for 8 visits with levocetrizine tablet 5mg on an on-demand basis. Urticaria activity score (UAS) sheet was provided to record and return every 2 weeks. Statistical analysis was done using the IBM SPSS 26 software package. Autologous serum skin test (ASST) was positive in 63% patients. Both the ASST positive and ASST negative group showed significant reduction in UAS7 score at week 14 compared to baseline. The reduction in mean UAS7 score was associated with a decreased pill burden and positive response in the patient and physician global assessment scale. No statistically significant difference between the two groups in terms of mean UAS7 reduction was found. This study has explored the efficacy and safety of autologous serum therapy in the pediatric CSU patients. Both ASST positive and ASST negative group respond to AST therapy.

Relevance of serum vascular endothelial growth factor (VEGF) and serum interleukin-10 in the severity of psoriasis in South Indian patients: A case–control study

Background: Psoriasis is a chronic inflammatory disorder and is associated with obesity, diabetes mellitus, and hypertension. There is an increased expression of inflammatory cytokines (interleukin [IL]-17, tumor necrosis factor [TNF]-α, IL-22, vascular endothelial growth factor [VEGF]) in the serum of psoriasis patients. Serum levels of IL-10, another anti-inflammatory cytokine, have been found at varying values in psoriasis in different regions of the world. Aims and Objectives: The aim of this article is to assess the serum IL-10 and serum VEGF in psoriasis patients with no co-morbidities and healthy controls. Materials and Methods: This study was conducted on 46 serum samples (23 psoriasis subjects and 23 healthy controls). After informed consent, 3 mL of serum was obtained and stored at -70°C. The samples were quantitatively assessed for VEGF-A and IL-10 by the enzyme-linked immunosorbent assay. Results: This study revealed that the mean (±SD) value of serum VEGF in cases was significantly higher than that in controls (cases = 235.21 ± 138.71; controls = 104.73 ± 36.01 pg/mL). However, levels of serum IL-10, although increased in cases (2.37 ± 1.61 pg/mL) when compared with controls (1.64 ± 0.89 pg/mL), showed no statistical significance. Conclusion: In this study, serum VEGF and IL-10 levels were increased in psoriasis when compared with controls but were not significantly related to the Psoriasis Area and Severity Index. The significant correlation between serum VEGF and IL-10 levels in cases when compared with controls suggests their role in the pathogenesis of psoriasis. Persistently increased values in psoriasis patients may lead to the development of comorbidities.

Reduced levels of serum EPA and DHA identified in patients with non-small-cell lung cancer using a new rapid validated LC-MS/MS method.

Eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) have been suggested to play roles in various diseases, yet there is little data on their changes in patients with non-small-cell lung cancer (NSCLC). A simple LC-MS/MS method for EPA and DHA determination is critical to exploring EPA and DHA level changes in NSCLC patients. 25 µL of serum was mixed with 25 µL of internal standard working solution, and then 450 µL of acetonitrile for protein precipitation. After vortex and centrifugation, the supernatant was directly used for LC-MS/MS analysis. The method was well validated with linearity, precision, recovery, and matrix effect. The concentrations of EPA and DHA in serum samples from 211 NSCLC patients and 227 healthy controls were determined by this LC-MS/MS method. Good separation and reliable quantification of EPA and DHA in serum samples were achieved by our method. Compared with healthy controls, serum EPA and DHA were significantly reduced in both adenocarcinoma and squamous cell carcinoma patients. The concentrations of EPA and DHA showed a progressive decrease in healthy controls, early- and advanced-stage NSCLC patients. This study identified significant reductions in serum EPA and DHA in NSCLC patients through the development of an LC-MS/MS method.

High-Throughput and In-Depth Proteomic Profiling of 5 μL Plasma and Serum Using TMTpro 16-Plex.

High-throughput and in-depth proteomic analysis of plasma and serum samples remains challenging due to the presence of multiple high-abundance proteins. Here, we provide a detailed protocol for proteomic analysis of serum and plasma specimens using a high-abundance protein depletion kit and TMTpro 16-plex reagents. This method requires only 5μL serum or plasma, identifying and quantifying about 1000 proteins. A batch of 16 samples can be processed in 36h. On average, each sample consumes about 1.5h of mass spectrometer instrument time. Overall, our method can identify proteins across six orders of magnitude with high reproducibility (CV<20%) using a shorter instrument time and less sample volume compared to existing methods. Thus, the method is suitable to be applied to large-scale proteomic studies.

Rapid Derivatization of Phenolic and Oxime Hydroxyl with Isonicotinoyl Chloride under Aqueous Conditions and Its Application in LC–MS/MS Profiling Multiclass Steroids

Quantification of steroids possesses a crucial clinical value in early diagnosis and prognosis evaluation of various endocrine diseases. However, it is still challenging to realize feasible analysis of estrogens, androgens, progestogens, and corticoids within one single workflow. In this study, two derivatization reactions were newly designed for improvement: (1) acylation of phenolic hydroxyl on estrogens with isonicotinoyl chloride (INC) under the catalysis of 4-dimethylaminopyridine and (2) post-modification of oxime hydroxyl on hydroxylamine-pretreated ketosteroids with INC. Both reactions could conduct instantaneously at room temperature under aqueous conditions. Moreover, the resulting phenolic-INC and oxime-INC esters exhibited favorable MS responses. Through integrating these derivatization strategies with cold-induced phase separation technology, a feasible LC-MS/MS method was developed for simultaneous quantification of 15 multiclass steroids with proper sample consumption (50 μL serum), satisfying sensitivity (lower limit of quantitation at 0.01-5.00 ng/mL) and high throughput (40 min for sample-preparation). The practical applicability was tested by detecting 30 real samples from pregnant and non-pregnant women. The obtained results showed a good agreement with a previous validated methodology.

Serum amino acids quantification by plasmonic colloidosome-coupled MALDI-TOF MS for triple-negative breast cancer diagnosis.

Triple-negative breast cancer (TNBC) is characterized with high diffusion, metastasis and recurrence. The early treatment and early diagnosis of TNBC are highly important for the survival of TNBC patients. Nevertheless, traditional methods for TNBC diagnosis, i.e. imaging examinations and tissue biopsy, cannot achieve early diagnosis, are invasive and associated with the risk of arousing tumor spreading. Herein, we developed colloidosomes-coupled matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) to quantify free serum amino acids for the diagnosis of TNBC. Gold nanoparticles (AuNPs) were used to form water-in-oil colloidosomes and to encapsulate deproteinated serum for MALDI-TOF MS analysis. With small sample spot size (200-300 μm) and densely packed AuNPs monolayer, the dried sample spot from colloidosomes can facilitate MALDI-TOF MS analysis of small molecule metabolites with high sensitivity, high reproducibility and good quantification performance. We used the method to quantify free amino acids in human serum. A cohort of 30 TNBC patients, 30 breast lump patients and 30 healthy controls were recruited for the study. It was found that the concentrations of free amino acids in TNBC patients were significantly lower than that of heathy controls, and the concentrations were also significantly different from that of breast lump patients. Based on the quantities of serum free amino acids, we have built a machine learning-based classification model to differentiate TNBC patients from the controls, including healthy controls and breast lump patients, and the sensitivity, specificity and accuracy were 95%, 100% and 97%, respectively. The assay consumes less than 1​μL serum per analysis, and takes only minutes to analyze a sample. With the advantages of low cost, low sample consumption, high throughput in analysis and high accuracy in identification, the non-invasive liquid biopsy method is promising to be applied to clinical diagnosis of TNBC.

An Alloy Platform of Dual‐Fingerprints for High‐Performance Stroke Screening

AbstractA multi‐modal serum profiling platform holds promise for precision diagnosis of diseases. Still, advanced tools are in demand to deliver the multi‐modal serum profiling. Herein, a bimodal spectrometric protocol is designed for stoke serum profiling using an alloy platform, by integrating label‐free surface‐enhanced Raman spectroscopy (SERS) and laser desorption/ionization mass spectrometry (LDI‐MS). The PdAu@Au concave cube with a wide localized surface plasmonic resonance (LSPR) range simultaneously enhances the signals from SERS and LDI‐MS, enabling high‐throughput co‐detection of vibrational and metabolic fingerprints of 0.1 µL serum in 2 min with simple pretreatment. Further, a dual‐fingerprints screening model of stroke is constructed, by adaptive machine learning with a programming nonlinear fitting model. The area under the curve are 0.949 (0.917–0.977, 95% confidence interval (CI)) and 0.911 (0.812–0.984, 95% CI), in the discovery and validation cohorts, respectively. Finally, five metabolites are identified that correlated to SERS signals and mapped the relevant pathways. This study features high performance in terms of throughput, speed, sample volume, and accuracy, providing new insight into the construction of multiplexed characterization platforms for precision diagnostic.

PepSeq: a fully in vitro platform for highly multiplexed serology using customizable DNA-barcoded peptide libraries.

PepSeq is an in vitro platform for building and conducting highly multiplexed proteomic assays against customizable targets by using DNA-barcoded peptides. Starting with a pool of DNA oligonucleotides encoding peptides of interest, this protocol outlines a fully in vitro and massively parallel procedure for synthesizing the encoded peptides and covalently linking each to a corresponding cDNA tag. The resulting libraries of peptide/DNA conjugates can be used for highly multiplexed assays that leverage high-throughput sequencing to profile the binding or enzymatic specificities of proteins of interest. Here, we describe the implementation of PepSeq for fast and cost-effective epitope-level analysis of antibody reactivity across hundreds of thousands of peptides from <1 µl of serum or plasma input. This protocol includes the design of the DNA oligonucleotide library, synthesis of DNA-barcoded peptide constructs, binding of constructs to sample, preparation for sequencing and data analysis. Implemented in this way, PepSeq can be used for a number of applications, including fine-scale mapping of antibody epitopes and determining a subject's pathogen exposure history. The protocol is divided into two main sections: (i) design and synthesis of DNA-barcoded peptide libraries and (ii) use of libraries for highly multiplexed serology. Once oligonucleotide templates are in hand, library synthesis takes 1-2 weeks and can provide enough material for hundreds to thousands of assays. Serological assays can be conducted in 96-well plates and generate sequencing data within a further ~4 d. A suite of software tools, including the PepSIRF package, are made available to facilitate the design of PepSeq libraries and analysis of assay data.

Sample multiplexing for increasing throughput for quantification of estrogens in serum by LC-MS/MS.

Estrogens are involved in many physiological processes in vivo. The accurate and rapid quantification of estrogens is required for the diagnosis and prognosis of estrogen-related diseases. To achieve high-volume assays, we developed and validated a sample-multiplexing liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the quantification of serum estrogens including estrone (E1), estradiol (E2), and estriol (E3). A total of 100 μL serum samples were extracted using ethyl acetate. After derivatization with either dansyl chloride or pyridine-3-sulfonyl chloride, derivatized samples were combined. Then we performed the second liquid-liquid extraction using hexane to purify the mixture. Finally, the reconstitution solutions were injected into LC-MS/MS. In addition, the proposed LC-MS/MS method was validated according to FDA and CLSI guidelines. Within a single run (7min), this sample-multiplexing LC-MS/MS method could simultaneously analyze E1, E2, and E3 in 2 serum samples. Meanwhile, the method demonstrated satisfactory analytical characteristics including accuracy (87.7-110.3%), linearity (2-1000pg/mL, R2 > 0.99), precision (intra-assay CV, 1.7-8.7%; inter-assay CV, 1.9-9.4%), and negligible interference and carry-over effect as well as acceptable matrix effect. In conclusion, this sample-multiplexing LC-MS/MS method has achieved a doubled-throughput assay for simultaneous quantification of E1, E2, and E3 without compromising analytical characteristics.

Electrophoretic detection of black market myostatin propeptide.

Myostatin propeptide is prohibited according to chapter S4 of the "WADA 2022 List of Prohibited Substances and Methods." So far, no approved myostatin-propeptide pharmaceuticals are available. Nevertheless, myostatin-propeptides can be bought on the black market for "research purposes." A study on black market myostatin propeptide products is presented as well as electrophoretic detection methods for serum and urine. Out of the 12 tested products, only nine actually contained the protein. Separation by SDS-PAGE revealed that the nine products were relatively impure and that the main compound had a much higher mass (approximately 54-55 kDa) than expected (approximately 33 kDa). Further analyses by mass spectrometry showed that the elevated molecular mass was due to the presence of a full length GST-tag on the propeptide. The developed detection method for serum is based on immunoprecipitation (IP) followed by SDS-PAGE and Western blotting. In total, three anti-myostatin propeptide antibodies were tested. All of them were well suited for either IP or immunoblotting. The final protocol applies a biotinylated polyclonal antibody, streptavidin-coated magnetic beads, and a monoclonal detection antibody. For a sample volume of 500 μL serum, the detection limit of the method is approximately 2.5 ng/mL. The urine method applies a commercial ELISA for IP and performs with a limit of detection (LOD) of approximately 0.4 ng/mL. Furthermore, practically all currently available myostatin propeptide standards were also investigated. Due to the significant molecular mass difference of the black market products, an unambiguous differentiation from endogenous myostatin propeptide is possible.

Simultaneous estimation of total homocysteine and methylmalonic-acid using LC-MS/MS: Clinical application in adult and pediatric

Quantitative determination of biomarkers homocysteine (Hcy) and methylmalonic acid (MMA), the regulators of cobalamin (Cbl) and folate levels, together used as a biomarkers to diagnose chemical insufficiency/deficiency of Cbl and folate. We report simultaneous clinical estimation of total Hcy and MMA with efficient clean-up, sensitive and selective LC-MS/MS method. Efficient sample clean-up was achieved by a two-step extraction protocol with 100 μL serum. The validated method was applied to 893 clinical samples from 2 cohorts including pediatrics and mothers, respectively, for identifying their Cbl and folate status. The method shows excellent order of linearity for Hcy (22.2nM-3.7 μM) and MMA (42.34 nM - 5.92 μM), respectively. Complete method validation was performed where intraday-interday accuracy-precision and mean stability recovery data were found within ±15%. The validated method was extended for the quantification of serum total Hcy-MMA levels in clinical samples. The efficient extraction with negligible matrix-effect (ME) has reduced LC-MS/MS chocking and clean-up downtime. The rapid, sensitive and robust LC-MS/MS method has been successfully validated for simultaneous estimation of total Hcy and MMA using only 100 μL serum. The method was applicable to large number of clinical samples and was found to be good throughput with low contamination of mass detector, high sensitivity and selectivity.

The Role of Interleukin 12 in Iraqi Patients with Psoriasis

Psoriasis is a chronic skin condition with immune-mediated inflammatory etiology. Many studies have indicated that Interleukin-12 (IL-12) plays an essential role in the severity of psoriasis.So to elucidate IL-12 role in regulating psoriasis promotion, the sera levels of IL-12 are detected in psoriasis patients and healthy people. The study contains 45 individuals with psoriasis, and the same number of healthy persons are included as controls .The PASI (Psoriasis Area and Severity Index) was used to assess the severity of psoriasis. Patients were divided into three groups depending on severity: mild psoriatic patients (13), moderate psoriatic patients(15), and severe psoriatic patients (17).Five ml of serum was obtained from each individual in per groups to determine IL-12 levels. The data demonstrated that the age range (36-45) has the highest prevalence of psoriatic patients (51.11 %). In addition, the female to male ratio in patients with psoriasis was 1.2:1.IL-12 levels in psoriasis patients were significantly higher than in healthy persons (3.55 pg/ml and 2.79 pg/ml). Furthermore, there was a significant association between blood IL-12 levels and PASI for psoriatic patient groups. The findings suggest that IL-12 has a key role in the psoriasis pathogenesis and that it might be utilized to evaluate psoriasis and as a follow-up indicator for those with it.

Effects of probiotics and high fructose diet on De Ritis ratio, TGF-B levels, and liver histopathology of Sprague Dawley rats

Background: Non-alcoholic fatty liver disease (NAFLD) is the most common chronic liver disease. Histopathological diagnosis has been the gold standard for diagnosing and determining the severity of non-alcoholic steatohepatitis (NASH). However, this method is invasive, especially for diagnosing NASH in children. This research aimed to study the effect of probiotics on the De Ritis ratio, TGF-β levels, and histopathological changes in rats given High Fat, High Fructose (HFHFr) diet to look at the correlation between the De Ritis ratio and liver histopathology in diagnosing NAFLD. Method: Twenty-one male Sprague Dawley rats were included and divided into three groups. In the first group, as control only received a normal chow diet for eight weeks. The second group received an HFHFr diet for eight weeks, and the last group received HFHFr for eight weeks and a combination of HFHFr and probiotics for another eight weeks. After decapitation, we took 3 ml of serum from each rat to measure AST, ALT, and TGF-β. A 4 μm thick liver tissue slide was taken and stained with Hematoxylin – Eosin Stain for histopathological analysis. We used the Non-Alcoholic Steatohepatitis (NAS) Score for measuring liver damage progression. Results: De Ritis ratio and TGF-β level did not significantly differ between probiotic and non-probiotic groups (p = 0.064 and 0.383), but there was a significant NAS score difference (p = 0.001) in probiotic and non-probiotic groups. This was followed by a significant correlation between the De Ritis ratio and the NAS score (r = 0.613, p = 0.003). Conclusion: Probiotic supplementation alleviated liver damage caused by the HFHFr diet but did not successfully reduce the De Ritis ratio or improve TGF-β.

95P Serum-based colorectal cancer detection using orphan noncoding RNAs

Early detection of colorectal cancer (CRC) can drastically improve survival odds, reduce treatment complexity and side effects, and improve patient quality of life. In this study we develop and evaluate an assay that uses 1 mL of serum for detection of CRC using orphan noncoding RNAs (oncRNAs) across different cancer stages.