Body Fluids Of Patients Research Articles

Uremic Toxin Receptor AhR Facilitates Renal Senescence and Fibrosis via Suppressing Mitochondrial Biogenesis.

Retention of metabolic end-products in the bodily fluids of patients with chronic kidney disease (CKD) may lead to uremia. The uremic toxin indoxyl sulfate (IS), a tryptophan metabolite, is an endogenous ligand of aryl hydrocarbon receptor (AhR). It is clarified that the upregulation and activation of AhR by IS in tubular epithelial cells (TECs) promote renal senescence and fibrosis. Renal TEC-specific knockout of AhR attenuates renal senescence and fibrosis, as well as the suppression of PGC1α-mediated mitochondrial biogenesis in ischemia reperfusion (IR)- or IS-treated CKD mice kidneys. Overexpression of peroxisome proliferator-activated receptor gamma coactivator 1-α (PGC1α) attenuates IS-induced cell senescence and extracellular matrix production in cultured TECs. Mechanistically, AhR is able to interact with PGC1α and promotes the ubiquitin degradation of PGC1α via its E3 ubiquitin ligase activity. In summary, the elevation and activation of AhR by the accumulated uremic toxins in the progression of CKD accelerate renal senescence and fibrosis by suppressing mitochondrial biogenesis via promoting ubiquitination and proteasomal degradation of PGC1α.

A review of the pleiotropic actions of the IFN-inducible CXC chemokine receptor 3 ligands in the synovial microenvironment.

Chemokines are pivotal players in instigation and perpetuation of synovitis through leukocytes egress from the blood circulation into the inflamed articulation. Multitudinous literature addressing the involvement of the dual-function interferon (IFN)-inducible chemokines CXCL9, CXCL10 and CXCL11 in diseases characterized by chronic inflammatory arthritis emphasizes the need for detangling their etiopathological relevance. Through interaction with their mutual receptor CXC chemokine receptor 3 (CXCR3), the chemokines CXCL9, CXCL10 and CXCL11 exert their hallmark function of coordinating directional trafficking of CD4+ TH1 cells, CD8+ T cells, NK cells and NKT cells towards inflammatory niches. Among other (patho)physiological processes including infection, cancer,and angiostasis, IFN-inducible CXCR3 ligands have been implicated in autoinflammatory and autoimmune diseases. This review presents a comprehensive overview of the abundant presence of IFN-induced CXCR3 ligands in bodily fluids of patients with inflammatory arthritis, the outcomes of their selective depletion in rodent models, and the attempts at developing candidate drugs targeting the CXCR3 chemokine system. We further propose that the involvement of the CXCR3 binding chemokines in synovitis and joint remodeling encompasses more than solely the directional ingress of CXCR3-expressing leukocytes. The pleotropic actions of the IFN-inducible CXCR3 ligands in the synovial niche reiteratively illustrate the extensive complexity of the CXCR3 chemokine network, which is based on the intercommunion of IFN-inducible CXCR3 ligands with distinct CXCR3 isoforms, enzymes, cytokines, and infiltrated and resident cells present in the inflamed joints.

Treatment of Crimean-Congo Haemorrhagic Fever by Favipiravir in a Patient with Novel Coronavirus Co-Infection.

Crimean-Congo haemorrhagic fever (CCHF) is a lethal zoonotic disease caused by an RNA virus that is a member of the Nairovirus genus in the Bunyaviridae family from the arbovirus group. CCHF is transmitted by Hyalomma ticks through direct contact with the blood and other bodily fluids of patients or infected animals. A 65-year-old man was admitted to the emergency unit with dry cough, myalgia and fever. He was treated with favipiravir. He had disseminated intravascular coagulopathy with thrombocytopenia in the setting of COVID-19 infection. He tested positive for both COVID-19 and CCHF. By the end of the fifth day of treatment, his laboratory parameters and clinical symptoms had normalized. Favipiravir is currently on the market for treating COVID-19 infection worldwide. It has also been used to treat CCHF in laboratory animals. To the best of our knowledge this is the first report of CCHF successfully treated with favipiravir, which could be a key drug for treating human CCHF. Clinicians should be alert for concomitant viral infections such as Crimean-Congo haemorrhagic fever, which share similar clinical and laboratory findings to COVID-19.The effectiveness of favipiravir for viral infections other than influenza and COVID-19, such as Crimean-Congo haemorrhagic fever, should be elucidated.

Concordance of epidermal growth factor receptor mutation detection in bodily fluids other than blood with tissue biopsy: A retrospective analysis

Background: The concept of liquid biopsy as a surrogate for tumor tissue is not only restricted to blood-derived samples but also includes other malignant bodily fluids such as cerebrospinal fluid, pleural fluid, pericardial fluid, and ascites. The application of liquid biopsy in patients with metastatic lung cancer, with disease progression on epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (TKI) treatment, is unexplored in India.Objective: We aimed to determine the presence of EGFR gene mutations in various bodily fluids of patients with EGFR-mutant progressive metastatic lung adenocarcinoma.Materials and Methods: In this retrospective study conducted at the Tata Memorial Hospital in Mumbai, India, data from 11 patients with advanced EGFR-mutant lung adenocarcinoma were analyzed. Between December 2017 and June 2018, the cell-free DNA (cfDNA) obtained from the malignant bodily fluids and plasma of 11 patients were tested for the presence of EGFR-sensitizing mutations using real-time reverse transcription-polymerase chain reaction. The status of EGFR-sensitizing mutations in plasma and other bodily fluids was compared with that of the baseline tissue biopsy. In addition, the status of the acquired T790M resistance mutation in exon 20 at disease progression was determined using plasma and other bodily fluids.Results: Of the 11 patients, at disease progression, EGFR sensitizing mutations (exon 19 deletion and exon 21 L858R insertion) could be detected in the plasma of 7 (63%) and in other bodily fluids of all the 11 (100%) patients. Thus, the concordance for EGFR-sensitizing mutation detection in the plasma and other bodily fluids with the baseline formalin-fixed paraffin-embedded tissue was 63% and 100%, respectively. In addition, the acquired T790M resistance mutation was detected in the plasma of 3 (27.3%) patients and other bodily fluids of 9 (81.8%) patients.Conclusion: The rate of detection of the T790M mutation is higher in the free DNA obtained from malignancy-involved bodily fluid supernatant as compared to the plasma cfDNA. Our results suggest that cfDNA obtained from bodily fluids other than plasma can be used for the detection of EGFR mutations and that the mutation status may be a useful predictor of the response to TKIs.

The effects of menopausal hormone therapy on proinflammatory cytokines and immunoglobulins in perimenopausal patients with type 2 diabetes mellitus and chronic obstructive pulmonary disease (COPD)

Aim. To determine the effects of menopausal hormone therapy dosage on levels of proinflammatory cytokines and immunoglobulins in bodily fluids of patients with type 2 diabetes mellitus (DM) and chronic obstructive pulmonary disease (COPD) during perimenopause. Materials and methods. The study included 119 perimenopausal females with moderate type 2 DM and stable COPD with signs of menopausal syndrome. Cytokine levels in bronchoalveolar lavage fluid and blood serum were measured with flow cytofluorometry (Вeckman Coulter FC500, USA) using a multiplex kit for human cytokines (BMS810FF) in adherence to the manufacturer’s instructions. The lower limit of quantification was 2.5-52.7 pg/mL. The following cytokines were studied: IL-1β, IL-6, IL-8, TNF-α and IF-γ. The data was analyzed with Flow CytomixProver 3.0 manufacturer-licensed software package. IgМ, IgG and IgА in the blood serum (Vektor-Best, Russia) were detected using an immunoenzymatic assay (Stat Fax 3200Analyzer, Awareness Technology, USA). Menopausal hormone therapy (MHT) with 17 beta-estradiol/dydrogesterone (standard - 2 mg, low - 1 mg, ultralow - 0.5 mg): 0.5/2.5 (estradiol 0.5 mg/dydrogesterone 2.5 mg) - ultralow-dose MHT, 1/10 (estradiol 1 mg/dydrogesterone 10 mg) - low-dose MHT; 2/10 (estradiol 2 mg/dydrogesterone 10 mg) - standard-dose MHT. Саrbohydrate metabolism was assessed in three groups. Results and discussion. While 2/10 MHT yielded the most prominent decrease in IL-1β, IL-6, IL-8, TNF-α and IF-γ levels in bronchoalveolar lavage fluid and blood serum, the effect was statistically insignificant compared to 0.5/2.5 and 1/10 MHT. Initially decreased IgM, IgG and IgA levels were elevated in all the three dosage groups with no significant differences between them. As to carbohydrate metabolism target values of glycemia were achieved in all three groups taking MHT. Conclusion. Standard-, low - and ultralow-dose MHT has positive effects on levels of proinflammatory cytokines and immunoglobulins characteristic of the association between type 2 DM, COPD and menopausal syndrome. The differences between the three dosage groups were statistically insignificant. Different dosage of MHT with dydrogesterone provide for improving impaired carbohydrate metabolism.

The effects of menopausal hormone therapy on proinflammatory cytokines and immunoglobulins in perimenopausal patients with type 2 diabetes mellitus and chronic obstructive pulmonary disease (COPD).

To determine the effects of menopausal hormone therapy dosage on levels of proinflammatory cytokines and immunoglobulins in bodily fluids of patients with type 2 diabetes mellitus (DM) and chronic obstructive pulmonary disease (COPD) during perimenopause. The study included 119 perimenopausal females with moderate type 2 DM and stable COPD with signs of menopausal syndrome. Cytokine levels in bronchoalveolar lavage fluid and blood serum were measured with flow cytofluorometry (Вeckman Coulter FC500, USA) using a multiplex kit for human cytokines (BMS810FF) in adherence to the manufacturer's instructions. The lower limit of quantification was 2.5-52.7 pg/mL. The following cytokines were studied: IL-1β, IL-6, IL-8, TNF-α and IF-γ. The data was analyzed with Flow CytomixProver 3.0 manufacturer-licensed software package. IgМ, IgG and IgА in the blood serum (Vektor-Best, Russia) were detected using an immunoenzymatic assay (Stat Fax 3200Analyzer, Awareness Technology, USA). Menopausal hormone therapy (MHT) with 17 beta-estradiol/dydrogesterone (standard - 2 mg, low - 1 mg, ultralow - 0.5 mg): 0.5/2.5 (estradiol 0.5 mg/dydrogesterone 2.5 mg) - ultralow-dose MHT, 1/10 (estradiol 1 mg/dydrogesterone 10 mg) - low-dose MHT; 2/10 (estradiol 2 mg/dydrogesterone 10 mg) - standard-dose MHT. Саrbohydrate metabolism was assessed in three groups. While 2/10 MHT yielded the most prominent decrease in IL-1β, IL-6, IL-8, TNF-α and IF-γ levels in bronchoalveolar lavage fluid and blood serum, the effect was statistically insignificant compared to 0.5/2.5 and 1/10 MHT. Initially decreased IgM, IgG and IgA levels were elevated in all the three dosage groups with no significant differences between them. As to carbohydrate metabolism target values of glycemia were achieved in all three groups taking MHT. Standard-, low- and ultralow-dose MHT has positive effects on levels of proinflammatory cytokines and immunoglobulins characteristic of the association between type 2 DM, COPD and menopausal syndrome. The differences between the three dosage groups were statistically insignificant. Different dosage of MHT with dydrogesterone provide for improving impaired carbohydrate metabolism.

The effect of initial periodontal treatment on plasma, gingival crevicular fluid and salivary levels of 8-hydroxy-deoxyguanosine in obesity

ObjectiveRecent studies have shown adverse effects on the periodontium from the increased production of reactive oxygen species (ROS) in obesity. The purpose of this study was to investigate the effects of obesity on 8-hydroxy-deoxyguanosine (8-OHdG) levels in the bodily fluids of patients with and without periodontal disease and to evaluate changes after initial periodontal treatment. DesignForty-five obese individuals and 45 normal-weight individuals were included in this study. Obese and normal-weight groups were classified into three sub-groups: chronic periodontitis (CP), gingivitis (G) and periodontally healthy controls (CTRL). Gingival crevicular fluid (GCF), plasma, saliva samples and clinical measurements were obtained at baseline and a month after initial periodontal treatment. Levels of 8-OHdG were analysed by ELISA. ResultsWhile plasma 8-OHdG levels were significantly higher at baseline in the obese patients with periodontal disease than in the normal-weight individuals (P<0.05), no significant differences in GCF and saliva 8-OHdG levels were found (P˃0.05). GCF and salivary 8-OHdG levels in obese patients with G and CP were significantly higher than in CTRL groups at baseline (P<0.05). After treatment, 8-OHdG levels were decreased in all groups with periodontal disease (P<0.01). Statistically significant positive correlations were observed between GCF 8-OHdG levels and GI in all the groups (P<0.001). ConclusionsThe significant increase of plasma 8-OHdG levels in obese patients did not correlate with saliva and GCF 8-OHdG levels when compared to normal-weight individuals. Periodontal treatment had a positive effect on the periodontal parameters and 8-OHdG levels of both obese and normal-weight individuals.

S-14 : Inflammasomes have extracellular activities that propagate inflammation

Microbes or endogenous danger signals trigger the assembly of cytosolic inflammasome receptors with the adapter ASC into speck-like aggregates. ASC specks recruit and activate caspase-1, which induces IL-1β cytokine maturation and pyroptotic cell death. We found that after inflammasome activation and pyroptotic cell death of activated macrophages fully assembled inflammasomes accumulated in the extracellular environment. Extracellular inflammasomes retained biological activity as they mediated further pro-caspase-1 and pro-IL-1β processing in the extracellular space. In addition, macrophages were found to phagocytose extracellular inflammasomes leading to lysosomal damage, activation of the NLRP3 inflammasome as well as nucleation of soluble ASC in the cytosol of cells that had phagocytosed inflammasomes. These processes resulted in caspase-1 activation and further propagation of IL-1β responses. Extracellular inflammasomes were also found in bodily fluids of patients with inflammatory diseases and in mouse models after experimental inflammasome activation. Moreover, anti-ASC Abs were identified in sera of patients and animals with autoimmunity. Anti-ASC Abs opsonized extracellular inflammasomes leading to further amplification of inflammatory responses in vitro and in vivo. Together, these findings reveal extracellular functions of inflammasomes that can act to perpetuate inflammatory responses.

Skin dose rate conversion factors after contamination with radiopharmaceuticals: influence of contamination area, epidermal thickness and percutaneous absorption

Skin contamination with radiopharmaceuticals can occur during biomedical research and daily nuclear medicine practice as a result of accidental spills, after contact with bodily fluids of patients or by inattentively touching contaminated materials. Skin dose assessment should be carried out by repeated quantification to map the course of the contamination together with the use of appropriate skin dose rate conversion factors. Contamination is generally characterised by local spots on the palmar surface of the hand and complete decontamination is difficult as a result of percutaneous absorption. This specific issue requires special consideration as to the skin dose rate conversion factors as a measure for the absorbed dose rate to the basal layer of the epidermis. In this work we used Monte Carlo simulations to study the influence of the contamination area, the epidermal thickness and the percutaneous absorption on the absorbed skin dose rate conversion factors for a set of 39 medical radionuclides. The results show that the absorbed dose to the basal layer of the epidermis can differ by up to two orders of magnitude from the operational quantity Hp(0.07) when using an appropriate epidermal thickness in combination with the effect of percutaneous absorption.

Quantitative GSTP1 hypermethylation in bodily fluids of patients with prostate cancer

Objectives To further determine the value of real-time quantitative methylation-specific polymerase chain reaction (MSP) of GSTP1 as a molecular tool for the detection of prostate adenocarcinoma. Recent studies have shown a high frequency (more than 90%) of GSTP1 gene promotor methylation in prostate adenocarcinoma and a lower frequency in DNA from serum and urine. Methods Tissue samples from 69 patients with early-stage prostate adenocarcinoma and 31 patients with benign prostatic hyperplasia were collected. Matched urine and plasma specimens were obtained preoperatively. After sodium-bisulfite treatment, extracted DNA was analyzed for GSTP1 promoter hypermethylation both by conventional and real-time quantitative MSP. Results In tissue samples, GSTP1 hypermethylation was detected in 63 (91.3%) of the 69 patients with cancer and in 9 (29%) of the 31 patients with benign prostatic hyperplasia. Conventional MSP detected GSTP1 hypermethylation in a larger number of urine and plasma samples than did real-time quantitative MSP (53.6% versus 31.9%, overall). In all positive bodily fluids, the paired tumor was also confirmed to be methylated. GSTP1 hypermethylation was detected by both MSP methods in only one urine sample (3.2%) from a patient with benign prostatic hyperplasia. Conclusions Although not quantitative, conventional MSP is currently more sensitive than real-time quantitative MSP in the detection of GSTP1 hypermethylation in bodily fluids from patients with prostate cancer with clinically localized disease. The value of quantitative determinations in monitoring and risk assessment remains to be further explored.

Differential diagnostic use of interleukin patterns in patients being monitored after transplantation

Background and aim: We investigated how far the determination of selected interleukins in bodily fluids of patients who had received kidney allografts can help to confirm the diagnosis of complications after transplantations. Materials and methods: Levels of soluble interleukin-2-receptor, interleukin 6 and interleukin 8 were determined in serum and urine of 79 patients. According to the type of diagnosis obtained with histological, serological and microbiological methods and to the clinical course, the groups “stable graft function without complication”, “allograft rejection”, “cytomegalovirus infection”, “systemic extrarenal bacterial infection”, “urinary tract infection” and “pyelonephritis” were created. Results and conclusions: The activation of the immune system in different ways depending on the trigger substance (alloantigen, virus, bacterium) and the possibility to differentiate systemic and local processes cause typical patterns of interleukin levels in serum and urine in conjunction with the above mentioned complications after kidney transplantation. Cytomegalovirus infections and systemic extrarenal bacterial infections differ from rejection by the unchanged urine interleukins IL 6 and IL 8, the local urinary tract infections differ from rejection by the unchanged serum interleukins. Acute pyelonephritis differs from rejection by the significantly higher serum IL 6 level. During our daily clinical work, the practical interleukin determinations were proven to be an important tool for early and differential diagnosis of complications after kidney transplantation.

Global Strategies for Disease Detection and Treatment: Proteomics

The field of proteomics holds great promise for identifying nontargeted, global molecular profiles “signatures” of normal and diseased cells. While genomics offers a wide array of tools for identifying mutated or dysregulated genes, proteomics offers the ability to measure post-translational modifications, protein stability, phosphorylation state, protein-protein interactions, and protein DNA-binding affinities. These biological events play major roles in the pathogenesis of disease and cannot be studied by DNA and mRNA efforts alone. Recent studies have shown that there is no specific correlation between mRNA abundance and protein expression levels in a cell at a given time. The discrepancy could arise from the control of mRNA translation, and the stability of mRNAs and proteins. Because almost all therapeutic intervention strategies or early detection technologies target expressed proteins, proteomic-based studies can provide fundamental information to characterize disease progression at the molecular level. Therefore, proteomics has particular relevance to diagnostics and to the identification of disease markers, as proteins can be assayed in serum and other biological fluids. Since the completion of human genome sequence, the discipline of proteomics has taken a center stage in defining the future of molecular diagnostics. This is due, to some extent, to the fact that the sequencing of the human genome and other important genomes has opened the door for proteomics by providing a sequence-based framework for mining the proteome. The growth in proteomic tools and proteomics is in an extraordinary growth phase. As a consequence, proteomics is witnessing an extraordinary growth in the number of new investigators and biotechnology companies that are taking an active interest in the field. Proteomics is evolving at a fast pace, as is evident from this special issue. Strategies for protein fractionation prior to analysis are increasingly being relied upon. Selective enrichment for a subset of proteins of interest can be achieved using a variety of techniques from centrifugation procedures to affinity capture. In addition to reducing sample complexity and increasing sensitivity, strategies based on the separate analysis of subcellular compartments provide the means to determine protein location in a cell. There is clearly interest in developing robust “industrial strength” proteomic platforms to achieve high-throughput and high sensitivity. During the lean years of proteomics, the field was largely dominated by a single approach, namely twodimensional (2-D) gels, backed up by an assortment of related tools, ranging from software for image analysis to mass spectrometry. Several technologies for protein profiling are emerging that are not 2-D gel based, including direct profiling by mass spectrometry and the use of protein microarrays. High throughput technologies, such as Surface-Enhanced Laser Desorption/Ionization Time-of-Flight (SELDI-TOF) and Matrix-Assisted Laser Desorption/Ionization Time-ofFlight (MALDI-TOF) technologies, are providing us with a non-biased, global discovery approach using patient serum, plasma, urine or other sources of secretions, to identify – in a single run – the expression patterns in thousands of small molecular mass proteins (< 20 kDa) based on their molecular mass and charge, offering a tool to determine which proteins are secreted from tumor cells or are measurable in the bodily fluids of patients. However, it is likely that no one technology platform will meet the needs of the wide spectrum of applications encompassed by proteomics. Diagnosing disease through proteomics may well require all the tools at our disposal to identify and validate the most promising markers.