MONITORING OF PROGESTERONE AND ESTRONE FECAL METABOLITES THROUGHOUT GESTATION IN EWES

Abstract The aim of this study was to monitor the progesterone and fecal estrone metabolites throughout gestation in ewes correlating with the serum levels of these steroid hormones. Therefore, fecal and serum samples were collected from 5 weeks before mating and gestation until two weeks postparturition. Serum levels of progesterone and estrone and their fecal metabolites were measured by enzyme immunoassay. Serum and fecal hormonal patterns showed a significant correlation for both hormones (R = 0.8572, P < 0.001 for progesterone and R = 0.5893, P < 0.001 for estrone). The fecal progesterone metabolite levels showed significant increasing values among the three thirds of pregnancies, consistent with the serum levels and with the literature. Additionally, the prepartum peak of estrone in the fecal matrix was identified but without observation in the serum matrix due to the blood collection interval used. Therefore, this study demonstrated the viability of progesterone and estrone monitoring throughout gestation using fecal samples, making noninvasive longitudinal endocrine monitoring throughout gestation possible in this species.

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Practical evaluation of two commercial immunoassays for the quantification of steroid hormones in human saliva

Journal of Cellular Biotechnology ◽

10.3233/jcb-209012 ◽

2020 ◽

pp. 1-9

Author(s):

Jonas Schmidt ◽

Folker Wenzel ◽

Frithjof Blessing

Keyword(s):

Steroid Hormones ◽

Steroid Hormone ◽

Serum Levels ◽

Human Saliva ◽

Clinical Diagnostics ◽

Serum Samples ◽

Large Variability ◽

Hormone Levels ◽

Practical Evaluation

BACKGROUND: There are certain issues using saliva as a sample for steroid hormone analysis in clinical diagnostics. Therefore, it is important to test whether currently available assays are suitable for this approach in a routine laboratory. OBJECTIVES: The analytical performance of two commercially available immunoassays was compared under routine conditions. In addition, salivary steroid hormone levels were compared with serum levels. METHODS: Saliva and serum samples were collected from healthy subjects. Afterwards, the concentration of cortisol, testosterone, progesterone and estradiol in saliva was measured by using the ADIVA Centaur ® XP platform and commercially available manual ELISA kits. Serum levels of the selected steroid hormones were determined by using the ADVIA Centaur ® platform. RESULTS: Currently, the ADVIA Centaur ® XP platform cannot be used for the quantification of steroid hormones in saliva due to an insufficient calibration range. Commercially available ELISA kits on the other side are well suitable for this application. CONCLUSIONS: There are commercially available immunoassays which are sensitive enough for the determination of steroid hormones from saliva and can be used under routine conditions. However, the interpretation of salivary steroid hormone levels stays a challenging task due to the large variability of the results.

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Significant Induction of Soluble TNFR2 Compared with TNFR1 in Serum Samples of HIV Patients with or without Antiretroviral Medication

Infectious Disorders - Drug Targets ◽

10.2174/1871526518666181016110409 ◽

2020 ◽

Vol 20 (2) ◽

pp. 175-181

Author(s):

Bismark Sarfo ◽

Zelalem Teka Haile ◽

Selase Deletsu ◽

Eric Akpanja Mensah ◽

Evelyn Yayra Bonney

Keyword(s):

Hiv Infection ◽

Serum Levels ◽

Antiretroviral Drugs ◽

Rank Test ◽

Blood Collection ◽

Serum Samples ◽

Hiv Patients ◽

Antiretroviral Medication ◽

Signed Rank Test ◽

The Difference

Background: Tumor necrosis factor and its receptors (sTNFR1 and sTNFR2) have been implicated in many infectious diseases. Identification of the key receptor (sTNFR1 or sTNFR2) which drives the immunopathogenesis of HIV infection is crucial in developing adjunctive therapy for HIV. Objective: This study determined the expression levels of sTNFR1 and sTNFR2 in antiretroviral therapy (ART) - experienced and naïve HIV patients. Methods: A total of 40 HIV patients comprising 30 with ART and 10 without ART were enrolled from the Pantang Hospital located in the Greater Accra Region of Ghana for data and blood collection. Serum concentrations of sTNFR1 and sTNFR2 were determined by ELISA. Mann- Whitney U test was used to examine differences in serum levels of sTNFR1 and sTNFR2 between patients on ART and ART naïve patients. Wilcoxon Signed-Rank test was performed to determine the difference between sTNFR1 and sTNFR2, and Kruskal Wallis test was conducted to compare the effect of different antiretroviral drugs on the levels of sTNFR1 and sTNFR2. P< 0.05 was considered statistically significant. Results: A Wilcoxon Signed-Ranks Test indicated serum levels of sTNFR2 was statistically significantly higher than sTNFR1 (Z=-5.51; p0.05 was considered0.001). Levels of sTNFR1 and sTNFR2 did not differ by ART status U =91.00 (Z = -1.84), p = 0.065 and U = 131.50 (Z = -0.58, p =0.560), respectively. There were not significant differences in levels of TNFR2 H(2) = 1.86, p=0.395 and sTNFR1 (H (2) = 4.37, p=0.113 across different ART combinations. Conclusion: Compared to sTNFR1, the level of sTNFR2 is significantly increased during HIV infection irrespective of ART status. The high sTNFR2 level is not associated with antiretroviral drugs and may be another potential target for therapeutic development. This is the first study of sTNFRs in African population.

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Autoantibodies to Thrombomodulin: Development of an Enzyme Immunoassay and a Survey of their Frequency in Patients with the Lupus Anticoagulant

Thrombosis and Haemostasis ◽

10.1055/s-0038-1648482 ◽

1992 ◽

Vol 67 (05) ◽

pp. 507-509 ◽

Cited By ~ 16

Author(s):

John Gibson ◽

Margaret Nelson ◽

Ross Brown ◽

Hatem Salem ◽

Harry Kronenberg

Keyword(s):

Enzyme Immunoassay ◽

Lupus Anticoagulant ◽

Specific Binding ◽

Technical Problem ◽

Serum Samples ◽

Citrate Buffer ◽

The Mean ◽

Sample Dilution ◽

Negative Population ◽

Serum Components

SummaryIn order to investigate the possibility that autoantibodies to thrombomodulin (TM) may exist in patients with the lupus anticoagulant (LA) and perhaps be implicated in the pathogenesis of recurrent thrombosis seen in such patients, we developed an enzyme-immunoassay to screen serum samples for anti-human TM activity. The major technical problem encountered in developing this assay was to reduce the non-specific binding of serum components from both the LA positive and the negative population. Considerable reduction of non-specific binding was achieved by use of a phosphate/citrate buffer at pH 8.0 and the use of an optimal sample dilution of 1/40. In addition, samples were always tested in parallel in blank wells and results are expressed as an OD ratio. Samples from 113 patients with the LA were assayed and compared to 78 patients referred for LA testing but found to be negative. The mean OD values for the LA positive patients (± SD) was 1.36 (0.44) with a range of 0.78-2.57. This was virtually identical to the values for the LA negative population (1.38 ± 0.40, range 0.76-2.77). The results of this study indicate that there is no evidence for the presence of a significant autoantibody activity to TM in patients with the LA when compared to LA negative patients. If such autoantibodies do exist their frequency must be quite low.

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Reassessing the Role of the Active TGF-β1 as a Biomarker in Systemic Sclerosis: Association of Serum Levels with Clinical Manifestations

Disease Markers ◽

10.1155/2016/6064830 ◽

2016 ◽

Vol 2016 ◽

pp. 1-6 ◽

Cited By ~ 13

Author(s):

Andréa Tavares Dantas ◽

Sayonara Maria Calado Gonçalves ◽

Anderson Rodrigues de Almeida ◽

Rafaela Silva Guimarães Gonçalves ◽

Maria Clara Pinheiro Duarte Sampaio ◽

...

Keyword(s):

Systemic Sclerosis ◽

Clinical Manifestations ◽

Serum Levels ◽

Vascular Involvement ◽

Digital Ulcers ◽

Serum Samples ◽

Peripheral Blood Mononuclear ◽

Pbmc Culture ◽

Significant Difference ◽

Skin Biopsies

Objective. To determine active TGF-β1 (aTGF-β1) levels in serum, skin, and peripheral blood mononuclear cell (PBMC) culture supernatants and to understand their associations with clinical parameters in systemic sclerosis (SSc) patients.Methods. We evaluated serum samples from 56 SSc patients and 24 healthy controls (HC). In 20 SSc patients, we quantified spontaneous or anti-CD3/CD28 stimulated production of aTGF-β1 by PBMC. The aTGF-β1 levels were measured by ELISA. Skin biopsies were obtained from 13 SSc patients and six HC, and TGFB1 expression was analyzed by RT-PCR.Results. TGF-β1 serum levels were significantly higher in SSc patients than in HC (p< 0.0001). Patients with increased TGF-β1 serum levels were more likely to have diffuse subset (p= 0.02), digital ulcers (p= 0.02), lung fibrosis (p< 0.0001), positive antitopoisomerase I (p= 0.03), and higher modified Rodnan score (p= 0.046). Most of our culture supernatant samples had undetectable levels of TGF-β1. No significant difference in TGFB1 expression was observed in the SSc skin compared with HC skin.Conclusion. Raised active TGF-β1 serum levels and their association with clinical manifestations in scleroderma patients suggest that this cytokine could be a marker of fibrotic and vascular involvement in SSc.

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Folate and Cobalamin Serum Levels in Healthy Children and Adolescents and Their Association with Age, Sex, BMI and Socioeconomic Status

Nutrients ◽

10.3390/nu13020546 ◽

2021 ◽

Vol 13 (2) ◽

pp. 546

Author(s):

Paulina Kreusler ◽

Mandy Vogel ◽

Anja Willenberg ◽

Ronny Baber ◽

Yvonne Dietz ◽

...

Keyword(s):

Socioeconomic Status ◽

Children And Adolescents ◽

Serum Levels ◽

Population Based ◽

Research Centre ◽

Healthy Children ◽

Serum Samples ◽

Lms Method ◽

Healthy Participants ◽

Age And Sex

This study proposes age- and sex-specific percentiles for serum cobalamin and folate, and analyzes the effects of sex, age, body mass index (BMI), and socioeconomic status (SES) on cobalamin and folate concentrations in healthy children and adolescents. In total, 4478 serum samples provided by healthy participants (2 months–18.0 years) in the LIFE (Leipzig Research Centre for Civilization Diseases) Child population-based cohort study between 2011 and 2015 were analyzed by electrochemiluminescence immunoassay (ECLIA). Continuous age-and sex-related percentiles (2.5th, 10th, 50th, 90th, 97.5th) were estimated, applying Cole’s LMS method. In both sexes, folate concentrations decreased continuously with age, whereas cobalamin concentration peaked between three and seven years of age and declined thereafter. Female sex was associated with higher concentrations of both vitamins in 13- to 18-year-olds and with higher folate levels in one- to five-year-olds. BMI was inversely correlated with concentrations of both vitamins, whilst SES positively affected folate but not cobalamin concentrations. To conclude, in the assessment of cobalamin and folate status, the age- and sex-dependent dynamic of the respective serum concentrations must be considered. While BMI is a determinant of both vitamin concentrations, SES is only associated with folate concentrations.

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AB0042 CYTOKINE NETWORK ELUCIDATED BY THE QUANTIFICATION OF MULTIPLE CYTOKINES IN THE SERUM SEQUENTIALLY SAMPLED FROM RA PATIENTS WHO WERE TREATED WITH BIOLOGIC DMARDS

Annals of the Rheumatic Diseases ◽

10.1136/annrheumdis-2020-eular.5109 ◽

2020 ◽

Vol 79 (Suppl 1) ◽

pp. 1323.2-1324

Author(s):

K. Sato ◽

S. Mamada ◽

C. Hayashi ◽

T. Nagashima ◽

S. Minota

Keyword(s):

Bone Erosion ◽

Feedback System ◽

Serum Levels ◽

Type I Interferons ◽

Type I ◽

Serum Samples ◽

Cytokine Network ◽

Biologic Dmards ◽

Type I Ifns ◽

Before And After

Background:Biologic disease modifying anti-rheumatic drugs (DMARDs) have demonstrated that proinflammatory cytokines such as interleukin (IL-) 6 and tumor necrosis factor (TNF) play important roles in the pathogenesis of rheumatoid arthritis (RA). Other cytokines, such as type I interferons (IFNs), are also implicated in its pathogenesis (ref 1). However, the complete picture of the cytokine network involved in RA remains to be elucidated.Objectives:By quantifying sets of cytokines in the serum of RA patients before and after treatment with various biologic DMARDs, we sought to determine the effects of drugs on (A) type I IFNs, (B) soluble IL-6 receptors, and (C) other cytokines.Methods:52 patients with RA were treated with various biologic DMARDs (tocilizumab (TOC): 16, abatacept (ABT): 15, and TNF inhibitors (TNFi): 21). Serum samples were obtained (1) before, (2) approximately 4 weeks after (3) and approximately 12 weeks after the initiation of treatment. A suspension bead-array system was used for analysis; Bio-Plex Human Cytokine 17-plex Assay kits and Express Custom Panels (Bio-Rad), including IFN-β, IFN-α2, soluble IL-6 receptor α (sIL6Rα) and gp130 were used.Results:(1) As expected, the disease activity score 28-joiny count (DAS28) using the erythrocyte sedimentation rate (ESR) significantly decreased in all three groups (TOC, ABT and TNFi) by 12 weeks.(2) IFN-α2 was barely detected in the serum samples. IFN-β seemed to increase slightly in the ABT group, but the increase was not statistically significant.(3) The levels of sIL6Rα did not change substantially. Those of gp130 decreased slightly but significantly in the TOC group by 12 weeks.(4) The levels of IL-6 decreased significantly in the ABT group by 12 weeks. Those in the TNFi group decreased significantly at 4 weeks but not 12 weeks (Fig. 1A).(5) The levels of IL-7 decreased significantly only in the TOC group (Fig. 1B).Conclusion:(1) The biologic DMARDs tested in this study did not significantly affect the serum levels of type I IFNs in this study.(2) The decrease in gp130 in the TOC group may imply that gp130 is induced by IL-6, although whether this level of decrease has physiological significance is open to question.(3) Serum IL-6 was significantly decreased in the TNFi group at 4 weeks but not 12 weeks. TNF has been reported to induce IL-6 (ref 2), but negative feedback loop(s) may be present. Such a feedback system might make the discontinuation of TNFi difficult, even if patients are in remission.(4) IL-7 may be a target of IL-6. A higher level of IL-7 has been reported to be present in the joints of RA patients compared with osteoarthrosis and it is a cytokine implicated in the differentiation of osteoclasts (ref 3). This may partly explain the effect of TOC on preventing bone erosion in RA.References:[1]Ann Rheum Dis. 2007; 66: 1008–14[2]Rheumatology 2007; 46: 920-6[3]Rheumatology 2008; 47: 753-9Acknowledgments:We thank all the members of the Division of Rheumatology and Clinical Immunology, Department of Medicine, Jichi Medical University. We are also grateful to the patients involved in this study.Disclosure of Interests:Kojiro Sato Grant/research support from: Abbie, Pfizer, Chugai, Astellas, Mitsubishi-Tanabe, Ono, Takeda, Sachiko Mamada: None declared, Chiyomi Hayashi: None declared, Takao Nagashima: None declared, Seiji Minota: None declared

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Dehydroepiandrosterone (DHEA) Serum Levels Indicate Cerebrospinal Fluid Levels of DHEA and Estradiol (E2) in Women at Term Pregnancy

Reproductive Sciences ◽

10.1007/s43032-021-00541-2 ◽

2021 ◽

Author(s):

Pardes Habib ◽

Joseph Neulen ◽

Shahin Habib ◽

Benjamin Rösing

Keyword(s):

Pregnant Women ◽

Strong Correlation ◽

Serum Levels ◽

Neuroactive Steroids ◽

Term Pregnancy ◽

Serum Samples ◽

Functional Changes ◽

The Central Nervous System ◽

Csf Levels ◽

Fluid Levels

AbstractNeuroactive steroids such as dehydroepiandrosterone (DHEA), estradiol (E2), and progesterone (P4) are associated with structural and functional changes in the central nervous system (CNS). Measurement of steroid levels in the CNS compartments is restricted in accessibility. Consequently, there is only limited human data on the distributional equilibrium for steroid levels between peripheral and central compartments. While some neuroactive steroids including DHEA and E2 have been reported to convey excitatory and proconvulsant properties, the opposite was demonstrated for P4. We aimed to elucidate the correlation between peripheral and central DHEA, E2, and P4 levels in women at term pregnancy. CSF and serum samples of 27 healthy pregnant women (22–39 years) at term pregnancy were collected simultaneously under combined spinal and epidural anesthesia and used for DHEA ELISA and E2, and P4 ECLIA. All three neuroactive steroids were detected at markedly lower levels in CSF compared to their corresponding serum concentrations (decrease, mean ± SD, 97.66 ± 0.83%). We found a strong correlation for DHEA between its serum and the corresponding CSF levels (r = 0.65, p = 0.003). Serum and CSF levels of E2 (r = 0.31, p = 0.12) appeared not to correlate in the investigated cohort. DHEA serum concentration correlated significantly with E2 (r = 0.58, p = 0.0016) in CSF. In addition, a strong correlation was found between DHEA and E2, both measured in CSF (r = 0.65, p = 0.0002). Peripheral DHEA levels might serve as an indicator for central nervous levels of the neuroactive steroids DHEA and E2 in pregnant women.

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PD-L1 expression in tumor lesions and soluble PD-L1 serum levels in patients with breast cancer: TNBC versus TPBC

Breast Disease ◽

10.3233/bd-201049 ◽

2021 ◽

pp. 1-9

Author(s):

Parvaneh Yazdanpanah ◽

Ali Alavianmehr ◽

Abbas Ghaderi ◽

Ahmad Monabati ◽

Mehdi Montazer ◽

...

Keyword(s):

Breast Cancer ◽

Tumor Cells ◽

Serum Levels ◽

Tumor Stage ◽

Serum Samples ◽

Healthy Women ◽

Grade 3 ◽

Anti Tumor Activity ◽

Tnbc Subtype ◽

Cell Death Protein

BACKGROUND: Block of programmed cell death protein 1 (PD-1) interaction with its ligand, PD-L1, enhances anti-tumor activity. OBJECTIVES: We aimed to assess the association between PD-L1 expression in tumor cells and CD8+ tumor infiltrating T cells (TILs) as well as soluble (s)PD-L1 serum levels in patients with triple negative breast cancer (TNBC) compared to triple positive (TPBC). METHODS: A total of 113 tumor sections and 133 serum samples were available from 144 patients with breast cancer (72 TNBC and 72 TPBC). Dual immunohistochemistry staining was applied to determine differential PD-L1 expression in tumor cells and CD8+ TILs. Soluble PD-L1 serum levels were also evaluated in patients compared to 40 healthy women by ELISA method. RESULTS: Despite TPBC patients which were mostly grades 1/2, TNBC patients were grade 3 (72% versus 66.7%, P < 0.001). Most of the TNBC patients were stages I/II, whereas most of the TPBC patients were stages III/IV (57.3% versus 68.3%,P = 0.005). There was no difference in tumor size and metastasis between TNBC and TPBC patients, although the number of involved lymph nodes was significantly more in TPBC patients (P = 0.0012). PD-L1 expression was detected in 11.5% of samples mostly in TNBC subtype and was associated with advanced grades (P = 0.039). There was no relationship between PD-L1 expression and tumor stage. PD-L1 expression in CD8+ TILs was nonsignificantly higher than tumor cells. Serum levels of sPD-L1 showed no difference between patients and healthy women. We found no correlation between PD-L1 expression in tumor lesions and serum levels of sPD-L1 in patients. CONCLUSION: PD-L1 expression was more detected in our patients with TNBC. It seems that, these patients who are resistant to standard chemotherapy regimens may get benefit from PD-L1 inhibition therapy and because of its low serum levels, sPD-L1 cannot interfere with this therapy.

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α-Tocopherol, lipids and lipoproteins in knee-joint synovial fluid and serum from patients with inflammatory joint disease

Clinical Science ◽

10.1042/cs0830657 ◽

1992 ◽

Vol 83 (6) ◽

pp. 657-664 ◽

Cited By ~ 51

Author(s):

K. Fairburn ◽

M. Grootveld ◽

R. J. Ward ◽

C. Abiuka ◽

M. Kus ◽

...

Keyword(s):

Synovial Fluid ◽

Low Density Lipoprotein ◽

Density Lipoprotein ◽

Serum Levels ◽

Biologically Active ◽

Joint Disease ◽

Inflammatory Joint Disease ◽

Serum Samples ◽

Control Serum ◽

Inflammatory Synovial Fluid

1. We have determined the antioxidant status of synovial fluid and serum of patients with inflammatory joint disease in terms of the biologically active lipid-soluble antioxidant, α-tocopherol. Synovial fluid concentrations of α-tocopherol were significantly lower relative to those of paired serum samples (P<0.001). Serum levels of α-tocopherol in these patients did not differ significantly from those in control serum. 2. Lower concentrations of cholesterol, triacylglycerol and low-density lipoprotein were also observed in patients' synovial fluid compared with matched serum samples. However, multiple regression analysis of the data indicated that there remained a significant depletion of α-tocopherol, which was largely independent of these co-variables, in inflammatory synovial fluid. These findings are consistent with the consumption of α-tocopherol within the inflamed joint via its role in terminating the process of lipid peroxidation. 3. Nuclear magnetic resonance spectroscopic analysis of matched inflammatory synovial fluid and serum confirmed lower concentrations of triacylglycerol in synovial fluid together with evidence of a shortened mean triacylglycerol chain length. The latter metabolic difference suggests an increased utilization of triacylglycerols for energy within the inflamed joint.

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Increased Migratory and Activation Cell Markers of Peripheral Blood Lymphocytes in an Experimental Model of Nephrotic Syndrome

Mediators of Inflammation ◽

10.1155/2015/209764 ◽

2015 ◽

Vol 2015 ◽

pp. 1-11 ◽

Cited By ~ 5

Author(s):

Wagner de Fátima Pereira ◽

Gustavo Eustáquio Alvim Brito-Melo ◽

Cláudia Martins Carneiro ◽

Dirceu de Sousa Melo ◽

Karine Beatriz Costa ◽

...

Keyword(s):

Nephrotic Syndrome ◽

Peripheral Blood ◽

Tissue Injury ◽

Serum Levels ◽

Peripheral Blood Lymphocytes ◽

Phenotypic Characterization ◽

Blood Collection ◽

Blood Leukocytes ◽

Male Adult

The present study aimed to evaluate the expression of CD80 and CD18 in subpopulations of peripheral blood leukocytes and oxidative kidney damage in rats with nephrotic syndrome (NS) induced by doxorubicin (Dox) in comparison to control animals at different time points. Male adult Wistar rats were submitted to 24-hour urine and blood collection for biochemical and immunological analysis at 7, 14, 21, and 28 days after Dox injection. After euthanasia, the kidneys were removed for histological analysis and the evaluation of oxidative stress. The phenotypic characterization of leukocytes was performed using flow cytometry. Dox-injected animals exhibited increased CD18 expression in cytotoxic T lymphocytes, NK cells, and monocytes and high CD80 expression in monocytes. Kidney oxidative damage was positively correlated with CD80 expression in monocytes and serum levels of creatinine. These results suggest that phagocytic and cytotoxic cells are preferentially recruited to the tissue injury site, which may contribute to kidney dysfunction in this animal model of NS. The blockade of integrin and costimulatory molecules may provide new therapeutic opportunities for NS.

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