An MRM-Based Multiplexed Quantification Assay for Human Adipokines and Apolipoproteins

Adipokines and apolipoproteins are key regulators and potential biomarkers in obesity and associated diseases and their quantitative assessment is crucial for functional analyses to understand disease mechanisms. Compared to routinely used ELISAs, multiple reaction monitoring (MRM)-based mass spectrometry allows multiplexing and detection of proteins for which antibodies are not available. Thus, we established an MRM method to quantify 9 adipokines and 10 apolipoproteins in human serum. We optimized sample preparation by depleting the two most abundant serum proteins for improved detectability of low abundant proteins. Intra-day and inter-day imprecision were below 16.5%, demonstrating a high accuracy. In 50 serum samples from participants with either normal weight or obesity, we quantified 8 adipokines and 10 apolipoproteins. Significantly different abundances were observed for five adipokines (adipsin, adiponectin, chemerin, leptin, vaspin) and four apolipoproteins (apo-B100/-C2/-C4/-D) between the body mass index (BMI) groups. Additionally, we applied our MRM assay to serum samples from normal weight children and human adipocyte cell culture supernatants to proof the feasibility for large cohort studies and distinct biological matrices. In summary, this multiplexed assay facilitated the investigation of relationships between adipokines or apolipoproteins and phenotypes or clinical parameters in large cohorts, which may contribute to disease prediction approaches in the future.

Download Full-text

Determinants of IGF1 and GH across the weight spectrum: from anorexia nervosa to obesity

Acta Endocrinologica ◽

10.1530/eje-10-0365 ◽

2010 ◽

Vol 163 (2) ◽

pp. 185-191 ◽

Cited By ~ 29

Author(s):

D J Brick ◽

A V Gerweck ◽

E Meenaghan ◽

E A Lawson ◽

M Misra ◽

...

Keyword(s):

Anorexia Nervosa ◽

Free Testosterone ◽

Cross Sectional Study ◽

Normal Weight ◽

The Body ◽

Obese Women ◽

Significant Determinant ◽

Serum Samples ◽

Cross Sectional ◽

Gh Secretion

ContextChronic starvation is characterized by GH resistance, and obesity is characterized by decreased GH secretion. In both extremes, IGF1 levels may be low and androgen levels may be abnormal.ObjectiveTo investigate the determinants of IGF1 and GH across the weight spectrum in women.DesignCross-sectional study.SettingClinical research center.Study participantsIn total, 32 women had participated in the study: 11 women with anorexia nervosa (AN), 11 normal-weight women, and 10 obese women of comparable mean age.InterventionNone.Main outcome measuresPooled hourly overnight serum samples assayed for IGF1, GH, estradiol (E2), testosterone, SHBG, insulin, free fatty acids, and trunk fat.ResultsFree testosterone was higher in obese women and lower in women with AN than in normal-weight women, and was the only independent (and positive) predictor of IGF1 levels, accounting for 14% of the variability (P=0.032) in the group as a whole. This relationship was stronger when obese women were excluded, with free testosterone accounting for 36% of the variability (P=0.003). Trunk fat accounted for 49% of the variability (P<0.0001) of GH, with an additional 7% of the variability attributable to E2 (P=0.042) in the group as a whole, but was not a significant determinant of GH secretion when obese women were excluded.ConclusionsFree testosterone is a significant determinant of IGF1 levels in women across the body weight spectrum. In contrast, GH secretion is differentially regulated at the extremes of the weight spectrum.

Download Full-text

‘The long tail of Covid-19’ - The detection of a prolonged inflammatory response after a SARS-CoV-2 infection in asymptomatic and mildly affected patients

F1000Research ◽

10.12688/f1000research.27287.2 ◽

2021 ◽

Vol 9 ◽

pp. 1349

Author(s):

Ivan Doykov ◽

Jenny Hällqvist ◽

Kimberly C. Gilmour ◽

Louis Grandjean ◽

Kevin Mills ◽

...

Keyword(s):

Inflammatory Response ◽

Drug Targets ◽

Multiplex Assay ◽

The Body ◽

Medical Complications ◽

Serum Samples ◽

Long Tail ◽

Disease Mechanisms ◽

Associated Proteins ◽

Inflammatory Processes

‘Long Covid’, or medical complications associated with post SARS-CoV-2 infection, is a significant post-viral complication that is being more and more commonly reported in patients. Therefore, there is an increasing need to understand the disease mechanisms, identify drug targets and inflammatory processes associated with a SARS-CoV-2 infection. To address this need, we created a targeted mass spectrometry based multiplexed panel of 96 immune response associated proteins. We applied the multiplex assay to a cohort of serum samples from asymptomatic and moderately affected patients. All patients had tested positive for a SARS-CoV-2 infection by PCR and were determined to be subsequently positive for antibodies. Even 40-60 days post-viral infection, we observed a significant remaining inflammatory response in all patients. Proteins that were still affected were associated with the anti-inflammatory response and mitochondrial stress. This indicates that biochemical and inflammatory pathways within the body can remain perturbed long after SARS-CoV-2 infections have subsided even in asymptomatic and moderately affected patients.

Download Full-text

Selective degradation of serum proteins is likely responsible for the spurious differences in innate immunity proteins observed in a type 1 diabetes study

F1000Research ◽

10.12688/f1000research.5384.1 ◽

2014 ◽

Vol 3 ◽

pp. 237

Author(s):

Wenbo Zhi ◽

Sharad Purohit ◽

Shan Bai ◽

Ashok Sharma ◽

Jin-Xiong She

Keyword(s):

Type 1 Diabetes ◽

Serum Proteins ◽

Multiple Reaction Monitoring ◽

Geographical Area ◽

Response To Therapy ◽

Control Group ◽

Experimental Medicine ◽

Serum Samples ◽

Sample Set

Discovery and validation of serum protein biomarkers is of vital importance for the prediction, mechanism elucidation and monitoring response to therapy of type 1 diabetes mellitus. In this study, we attempted to replicate the results published in a 2013 issue of The Journal of Experimental Medicine by Qibin Zhang and colleagues described the discovery, verification and validation of several serum proteins/peptides that were drastically different between type 1 diabetes (T1D) patients and healthy controls, using label-free quantitative LC-MS-based proteomics and a multiple reaction monitoring mass spectrometry (MRM-MS) based multiplexed peptide assays. We performed the same MRM-MS assay in a large sample panel of 145 T1D patients and 156 autoantibody negative (AbN) control subjects (PANDA sample set) collected in the same geographical area, during the same period of time and by the same investigators, as well as 144 serum samples from the original authors (DASP sample set). Our measurement of 12 transitions/peptides in the DASP samples correlated very nicely with the authors’ published results, indicating that the techniques used in the two laboratories yield very similar results on the same sample sets. Yet, in our PANDA samples, five of the twelve peptides (LLDSLPSDTR, FQPTLLTLPR, TNLESILSYPK, LVLLNAIYLSAK and ITQVLHFTK) whose serum levels are significantly different in the DASP verification and/or blind sample sets are not significant (p>0.05). Only one peptide (TGAQELLR) showed marginal significance (p=0.03). Although the remaining 6 peptides (NIQSLEVIGK, TLEAQLTPR, ELDESLQVAER, AGALNSNDAFVLK, TFTLLDPK and DIPTNSPELEETLTHTITK) are significantly different between the T1D and control group in our PANDA sample set, the inter-group differences as measured by fold change (FC) are very small (FC = 1.0±0.1). Therefore, our results do not support the major findings in the report.

Download Full-text

‘The long tail of Covid-19’ - The detection of a prolonged inflammatory response after a SARS-CoV-2 infection in asymptomatic and mildly affected patients

F1000Research ◽

10.12688/f1000research.27287.1 ◽

2020 ◽

Vol 9 ◽

pp. 1349

Author(s):

Ivan Doykov ◽

Jenny Hällqvist ◽

Kimberly C. Gilmour ◽

Louis Grandjean ◽

Kevin Mills ◽

...

Keyword(s):

Inflammatory Response ◽

Drug Targets ◽

Multiplex Assay ◽

The Body ◽

Medical Complications ◽

Serum Samples ◽

Long Tail ◽

Disease Mechanisms ◽

Associated Proteins ◽

Inflammatory Processes

Download Full-text

EFFECT OF BODY MASS INDEX ON PEAK EXPIRATORY FLOW RATE

International Journal of Medical and Biomedical Studies ◽

10.32553/ijmbs.v3i9.570 ◽

2019 ◽

Vol 3 (9) ◽

Author(s):

K. Subramanyam ◽

Dr. P. Subhash Babu

Keyword(s):

Body Mass Index ◽

Body Mass ◽

Peak Expiratory Flow ◽

Flow Rate ◽

Peak Expiratory Flow Rate ◽

Normal Weight ◽

The Body ◽

Health Care Centre ◽

Expiratory Flow ◽

Group B

Obesity has become one of the major health issues in India. WHO defines obesity as “A condition with excessive fat accumulation in the body to the extent that the health and wellbeing are adversely affected”. Obesity results from a complex interaction of genetic, behavioral, environmental and socioeconomic factors causing an imbalance in energy production and expenditure. Peak expiratory flow rate is the maximum rate of airflow that can be generated during forced expiratory manoeuvre starting from total lung capacity. The simplicity of the method is its main advantage. It is measured by using a standard Wright Peak Flow Meter or mini Wright Meter. The aim of the study is to see the effect of body mass index on Peak Expiratory Flow Rate values in young adults. The place of a study was done tertiary health care centre, in India for the period of 6 months. Study was performed on 80 subjects age group 20 -30 years, categorised as normal weight BMI =18.5 -24.99 kg/m2 and overweight BMI =25-29.99 kg/m2. There were 40 normal weight BMI (Group A) and 40 over weight BMI (Group B). BMI affects PEFR. Increase in BMI decreases PEFR. Early identification of risk individuals prior to the onset of disease is imperative in our developing country. Keywords: BMI, PEFR.

Download Full-text

STUDIES IN THE SUPPOSED GOITROGENOUS EFFECT OF TAP WATER IN HELSINKI

Acta Endocrinologica ◽

10.1530/acta.0.xxxiii0630 ◽

1960 ◽

Vol XXXIII (IV) ◽

pp. 630-636

Author(s):

F.-E. Krusius ◽

P. Peltola

Keyword(s):

Tap Water ◽

Physiological Saline ◽

Normal Weight ◽

The Body ◽

Iodine Uptake ◽

Redistilled Water ◽

Detectable Difference ◽

Term Administration ◽

Long Term Experiment

ABSTRACT The study reported here was performed in order to examine the tap water of Helsinki for its alleged goitrogenous effect. In a short-term, 24-hour experiment with rats, kept on an iodine-poor diet, we noticed no inhibition of the 4-hour 131I uptake, as compared with that of animals receiving physiological saline instead of tap water. Two similar groups of rats receiving 1 and 2 mg of mercazole in redistilled water showed a distinct blockage of the 4-hour uptake, which proved the effect of this substance. In a long-term experiment of 5 weeks' duration there was no detectable difference in the body weight, thyroid weight and the 4-hour 131I uptake when the rats receiving tap water or distilled water to which 0.45 per cent of sodium chloride was added were compared with each other. Replacement of tap water by a 10 mg per cent solution of mercazole in redistilled water enlarged the thyroid to double its normal weight and increased the 131I uptake to approximately five times that of the controls. Thus our experiments failed to demonstrate any goitrogenous effect in the tap water of Helsinki. Changes similar to those produced by a long-term administration of mercazole, i. e. an enlargement of the thyroid and an increased thyroidal iodine uptake, have been shown to be due to milk collected from goitrous areas. The observations here reported confirm the importance of milk in the genesis of the goitre endemia of Helsinki. Attention is further called to the fact that a thyroidal enlargement combined with an increased thyroidal iodine uptake cannot always be taken as a sign of iodine deficiency because similar changes may be produced by the administration of goitrogens.

Download Full-text

Computer-supported Detection of M-Components and Evaluation of Immunoglobulins after Capillary Electrophoresis

Clinical Chemistry ◽

10.1093/clinchem/47.1.110 ◽

2001 ◽

Vol 47 (1) ◽

pp. 110-117 ◽

Cited By ~ 16

Author(s):

Magnus Jonsson ◽

Joyce Carlson ◽

Jan-Olof Jeppsson ◽

Per Simonsson

Keyword(s):

Capillary Electrophoresis ◽

Decision Support ◽

Protein Separation ◽

Serum Proteins ◽

Cost Effective ◽

Clinical Samples ◽

Serum Samples ◽

Computerized Decision Support ◽

Cost Effective Method ◽

Mathematical Algorithms

Abstract Background: Electrophoresis of serum samples allows detection of monoclonal gammopathies indicative of multiple myeloma, Waldenström macroglobulinemia, monoclonal gammopathy of undetermined significance, and amyloidosis. Present methods of high-resolution agarose gel electrophoresis (HRAGE) and immunofixation electrophoresis (IFE) are manual and labor-intensive. Capillary zone electrophoresis (CZE) allows rapid automated protein separation and produces digital absorbance data, appropriate as input for a computerized decision support system. Methods: Using the Beckman Paragon CZE 2000 instrument, we analyzed 711 routine clinical samples, including 95 monoclonal components (MCs) and 9 cases of Bence Jones myeloma, in both the CZE and HRAGE systems. Mathematical algorithms developed for the detection of monoclonal immunoglobulins (MCs) in the γ- and β-regions of the electropherogram were tested on the entire material. Additional algorithms evaluating oligoclonality and polyclonal concentrations of immunoglobulins were also tested. Results: CZE electropherograms corresponded well with HRAGE. Only one IgG MC of 1 g/L, visible on HRAGE, was not visible after CZE. Algorithms detected 94 of 95 MCs (98.9%) and 100% of those visible after CZE. Of 607 samples lacking an MC on HRAGE, only 3 were identified by the algorithms (specificity, 99%). Algorithms evaluating total gammaglobulinemia and oligoclonality also identified several cases of Bence Jones myeloma. Conclusions: The use of capillary electrophoresis provides a modern, rapid, and cost-effective method of analyzing serum proteins. The additional option of computerized decision support, which provides rapid and standardized interpretations, should increase the clinical availability and usefulness of protein analyses in the future.

Download Full-text

Accuracy of Age-Based Formula to Predict the Size and Depth of Cuffed Oral Preformed Endotracheal Tubes in Children Undergoing Tonsillectomy

Ear Nose & Throat Journal ◽

10.1177/0145561320980511 ◽

2021 ◽

pp. 014556132098051

Author(s):

Matula Tareerath ◽

Peerachatra Mangmeesri

Keyword(s):

Body Mass Index ◽

Endotracheal Tube ◽

Body Mass ◽

Normal Weight ◽

Overweight And Obesity ◽

The Body ◽

Insertion Depth ◽

Endotracheal Tubes ◽

Endotracheal Tube Size ◽

Actual Size

Objectives: To retrospectively investigate the reliability of the age-based formula, year/4 + 3.5 mm in predicting size and year/2 + 12 cm in predicting insertion depth of preformed endotracheal tubes in children and correlate these data with the body mass index. Patients and Methods: Patients were classified into 4 groups according to their nutritional status: thinness, normal weight, overweight, and obesity; we then retrospectively compared the actual size of endotracheal tube and insertion depth to the predicting age-based formula and to the respective bend-to-tip distance of the used preformed tubes. Results: Altogether, 300 patients were included. The actual endotracheal tube size corresponded with the Motoyama formula (64.7%, 90% CI: 60.0-69.1), except for thin patients, where the calculated size was too large (0.5 mm). The insertion depth could be predicted within the range of the bend-to-tip distance and age-based formula in 85.0% (90% CI: 81.3-88.0) of patients. Conclusion: Prediction of the size of cuffed preformed endotracheal tubes using the formula of Motoyama was accurate in most patients, except in thin patients (body mass index < −2 SD). The insertion depth of the tubes was mostly in the range of the age-based-formula to the bend-to-tip distance.

Download Full-text

The Effect of Dietary Intake and Nutritional Status on Anthropometric Development and Systemic Inflammation: An Observational Study

International Journal of Environmental Research and Public Health ◽

10.3390/ijerph18115635 ◽

2021 ◽

Vol 18 (11) ◽

pp. 5635

Author(s):

Roxana Maria Martin-Hadmaș ◽

Ștefan Adrian Martin ◽

Adela Romonți ◽

Cristina Oana Mărginean

Keyword(s):

Serum Proteins ◽

Caloric Intake ◽

Saturated Fat ◽

The Body ◽

Total Serum ◽

Fat Intake ◽

Trans Fats ◽

Study Objective ◽

Inflammatory Status ◽

Total Fat

(1) Background: Daily caloric intake should aim to reduce the risk of obesity or poor anthropometric development. Our study objective was to analyze the association between food consumption, inflammatory status and anthropometric development; (2) Methods: We performed a prospective observational analytical research during September 2020 and April 2021 on a group of 160 healthy subjects, aged between 6 and 12 years old, by analyzing food ingestion, the basal metabolic rate, anthropometric development and the inflammatory status; (3) Results: IL-6 was significantly correlated to the sum of skinfolds, along with both serum proteins and triglycerides. The skin folds were significantly correlated with the caloric intake and with total fat intake, next to saturated and trans fats. Unlike the skin folds, the body weight was significantly correlated with the caloric intake along with some vitamins, such as Vitamin A and Vitamin B12. Inactive mass increased with excessive folic acid, Vitamin E, Vitamin K and saturated fat intake; (4) Conclusions: The inflammatory status was influenced by the ingestion of micronutrients, total serum lipids and proteins. The anthropometric development was associated with the ingestion of carbohydrates, energy balance and energy intake. We can conclude that daily menu and nutrition imbalances can influence both the risk of obesity and the inflammatory status.

Download Full-text

Association of body mass index and maternal age with first stage duration of labour

Scientific Reports ◽

10.1038/s41598-021-93217-5 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Louise Lundborg ◽

Xingrong Liu ◽

Katarina Åberg ◽

Anna Sandström ◽

Ellen L. Tilden ◽

...

Keyword(s):

Body Mass Index ◽

Body Mass ◽

Maternal Age ◽

Normal Weight ◽

The Body ◽

Younger Women ◽

Individualized Approach ◽

Quantile Regression Analysis ◽

Labour Time ◽

Group 1

AbstractTo evaluate associations between early-pregnancy body mass index (BMI) and active first stage labour duration, accounting for possible interaction with maternal age, we conducted a cohort study of women with spontaneous onset of labour allocated to Robson group 1. Quantile regression analysis was performed to estimate first stage labour duration between BMI categories in two maternal age subgroups (more and less than 30 years). Results show that obesity (BMI > 30) among younger women (< 30 years) increased the median labour duration of first stage by 30 min compared with normal weight women (BMI < 25), and time difference estimated at the 90th quantile was more than 1 h. Active first stage labour time differences between obese and normal weight women was modified by maternal age. In conclusion: (a) obesity is associated with longer duration of first stage of labour, and (b) maternal age is an effect modifier for this association. This novel finding of an effect modification between BMI and maternal age contributes to the body of evidence that supports a more individualized approach when describing labour duration.

Download Full-text