scholarly journals Recombinant Antibodies with Unique Specificities Allow for Sensitive and Specific Detection of Uncarboxylated Osteocalcin in Human Circulation

2020 ◽  
Vol 107 (6) ◽  
pp. 529-542
Author(s):  
Milja Arponen ◽  
Eeva-Christine Brockmann ◽  
Riku Kiviranta ◽  
Urpo Lamminmäki ◽  
Kaisa K. Ivaska

Abstract Osteocalcin is a bone-specific protein which contains three glutamic acid residues (Glu) that undergo post-translational gamma-carboxylation. Uncarboxylated osteocalcin (ucOC) may participate in the regulation of glucose metabolism, thus measurement of ucOC could be useful in evaluating interactions between bone and glucose metabolism. We developed recombinant antibodies and immunoassay to specifically detect ucOC in human blood samples. ucOC-specific recombinant antibodies were selected from an antibody library by phage display. Four candidates were characterized, and one (Fab-AP13) was used to set up an immunoassay with a pre-existing MAb. Plasma ucOC levels were measured in subjects with normal fasting blood glucose (≤ 6 mmol/l, N = 46) or with hyperglycemia (≥ 7 mmol/l, N = 29). Further, we analyzed ucOC in age- and gender-matched patients with diagnosed type 2 diabetes (T2D, N = 49). Antibodies recognized ucOC without cross-reaction to carboxylated osteocalcin. Antibodies had unique binding sites at the carboxylation region, with Glu17 included in all epitopes. Immunoassay was set up and characterized. Immunoassay detected ucOC in serum and plasma, with on average 1.6-fold higher levels in plasma. ucOC concentrations were significantly lower in subjects with hyperglycemia (median 0.58 ng/ml, p = 0.008) or with T2D diagnosis (0.68 ng/ml, p = 0.015) than in subjects with normal blood glucose (1.01 ng/ml). ucOC negatively correlated with fasting plasma glucose in subjects without T2D (r = − 0.24, p = 0.035) but not in T2D patients (p = 0.41). Our immunoassay, based on the novel recombinant antibody, allows for specific and sensitive detection of ucOC in human circulation. Correlation between ucOC and plasma glucose suggests interactions between osteocalcin and glucose metabolism in humans.

2020 ◽  
Vol 28 (2) ◽  
pp. 112-119
Author(s):  
Raveendra P Gadag ◽  
Puneeth S Nayak ◽  
Tejaswini J

Background: Hearing impairment is known to hamper the quality of life among patients, especially among diabetics due to the association of neuropathy with diabetes. However, the prevalence and degree of the SNHL depends upon different factors, such as age, gender, disease duration of DM, family history and glycemic status of the patients. Therefore, this study aimed to assess the association of SNHL with DM duration and familial DM and gender preponderance among SNHL–DM patients. Methods: Total 140 patients with DM were assessed for hearing impairment using Rinne, Weber and Absolute Bone Conduction Tests along with pure tone audiometry. Patients’ glycaemic status was determined by estimating fasting blood glucose (FBG) and post prandial blood glucose (PPBG) levels. Independent t-test, chi-square, ANOVA and Pearson’s correlation tests along with linear regression model were used to find association and correlation using R software. Results: Out of 140 patients, 60 were suffering from SNHL and majority were suffering from bilateral minimal hearing loss. SNHL was significantly associated with family history, age, duration of DM, FBG and PPBG levels were(Pvalues:1.79E08, 4.41E-06 and 0.02), however, significant correlated with duration of DM, FBS and PPBG level (r value:0.14–0.41).Furthermore, significant SNHL at 500 and 8000Hz was observed in the present study (Pvalue:0.002). Conclusion: A conclusive proof was drawn that family history of DM serve as a valuable variable in assessing the SNHL among DM patients.


2014 ◽  
Vol 2014 ◽  
pp. 1-5 ◽  
Author(s):  
Xiaofei Huang ◽  
Pan Weng ◽  
Huixin Zhang ◽  
Yingli Lu

Objective. As a complicated symbiotic system, intestinal flora is reported closely related to the development of type 2 diabetes recently. Sleeve gastrectomy is one of the approaches of bariatric surgery and could improve blood glucose control in type 2 diabetes patients. This study was to explore the relationship between remodeled intestinal flora and glucose metabolism in diabetic rats.Methods. 20 male diabetic rats were operated; 10 of them underwent sleeve gastrectomy, and 10 of them underwent sham operation. Meanwhile 10 male normal rats underwent sleeve gastrectomy as control. The animals’ weight and FBG had been measured. The composition changes of intestinal flora were detected by 16S rDNA sequence analysis.Results. In diabetic rats, weight and fasting blood glucose decreased significantly after sleeve gastrectomy. However, there was no significant change for weight and blood glucose in normal rats after operation. The intestinal flora of diabetic rats reduced in the proportion of Firmicutes and increased in the proportion of Bacteroidetes after sleeve gastrectomy.Conclusion. The change of dominant microorganisms in intestinal flora might play an important role in the glucose metabolism.


Blood ◽  
2018 ◽  
Vol 132 (Supplement 1) ◽  
pp. 2350-2350
Author(s):  
Zeynep Karakas ◽  
Cihangir Sevimli ◽  
Nurdan Gul ◽  
Rana Comert ◽  
Memduh Dursun

Abstract The relationship between pancreatic iron overload and glucose dysregulation has not been well defined and need further investigation. Pancreas iron load measurements are also necessary to understand how hepatic, pancreatic, and cardiac iron burden prospectively modulate the evolution of diabetes in transfusion dependent thalassemia (TDT). Aim: This study aims to evaluate the presence of pancreatic iron overload among TDT patients in Turkey based on the MRI R2* and T2* results as well as to determine its association to Fasting blood glucose (FBG), insulin, fructosamine, Homa-IR, C-peptide, ferritin, liver, cardiac and hypophysis MRI R2* results. Material and Methods: This study was conducted at the Thalassemia Center in Istanbul University, Istanbul Medical Faculty. Forty-one TDT patients included the study with a mean age of 29.9±9,9 (median 29) years old, ranging from 11 to 45 years old. Pancreatic, liver, cardiac and hypophsis MRI's were conducted using 1.5 Tesla Philips MRI machine in the Department of Radiology. Pancreatic MRI R2* results < 30 Hz were considered normal, 30-100 Hz as mild, and > 100 Hz as moderate to severe pancreatic siderosis. Pancreatic MRI T2 results >26 ms were is normal, < 10 ms is severe iron overload as presented at literature. Correlations between pancreatic MRI R2* results and FBG, insulin, fructosamine, Homa-IR, C-peptide, ferritin, cardiac and hypophysis MRI R2* were evaluated using the Pearson correlation, Man Whitney U test. Results: Laboratory findings of glucose metabolism and MRI's of liver, pancreas, hypophsis and cardiac iron are shown at Table 1. There were no significant relationship between fasting blood glucose, insülin, C-peptid level, fructosamine, Homa-IR, ferritin and Pancreas T2* and R2* results (p>0.05). There was significant correlation between liver and pancreas MRI T2* and R2* results. (p<0.01). There was no significant correlation between hypophysis global R2, cardiac R2*, Pancreas T2* and, Pancreas R2* measurements (p>0.05). Fourteen patients were found to have normal pancreatic MRI R2* (34,1%) while 19 patients have normal pancreatic MRI T2 (46.3%). It is reported that T2 is irrelevant to age and sex, and R2 can be related to age-related fatty degeneration. There was significant pancreatic iron burden in 31.7% of patients (R2*>100 Hz), of whom only 1 patient was under 18 years of age.There were 22 (53.6%) patients with T2* <26 ms and had iron overload, 13 (31.7%) had T2 <10 ms and had significant iron overload. Almost three-quarters of patients (n:13, 31.7%) had moderate to severe pancreatic iron load on both T2* and R2* measurements. Thirteen patients with significant pancreatic iron overload had 3 severe hypophysis, 1 moderate, 2 severe cardiac siderosis. and had no serious liver iron burden. This shows that the treatment of chelation first led to liver, then heart and pituitary, respectively, and finally pancreatic iron burden and demonstrates the importance of pancreas MR in assessing the body iron load. Discussion:The lack of correlation between pancreatic and cardiac MRI R2* are in contrast to another demonstrating that pancreatic iron load are good predictors of cardiac iron load. This is due to the fact that our patients are followed up with cardiac MR and taken to the intensive chelation programs of the patients who detect iron in the heart. Previous studies also report that while pancreatic R2* may be sensitive for glucose dysregulation, most patients may still have no symptoms and normal blood glucose, indicating the presence of a preclinical iron burden. Pancreatic MRI R2* can detect pancreatic iron accumulation at a much earlier stage. Lack of excessive iron load in liver of our patient can be explain low glucose metabolism disorder despite high pancreatic iron overload. Disclosures No relevant conflicts of interest to declare.


2019 ◽  
Vol 185 (3-4) ◽  
pp. 346-353
Author(s):  
Weniko Caré ◽  
Jessica Hénard ◽  
Jonathan Gillard ◽  
Serge Cremades ◽  
Franck Ceppa ◽  
...  

Abstract Introduction In the French armed forces, the biological checkup required during the recruitment process comprises a urinalysis (urinary dipstick), a complete blood count (CBC), and measurement of serum levels of aspartate aminotransferase, alanine aminotransferase, fasting blood glucose, and creatinine. This study aimed to evaluate the benefits of this biological checkup and to determine the most relevant parameters. Materials and Methods We conducted a monocentric retrospective study of all standardized and systematically conducted blood tests (CBC and measurement of aspartate aminotransferase, alanine aminotransferase, fasting blood glucose, and creatinine) over a 15-month period among 726 French Army recruits. Results The population included mainly young males (85.4%, mean age 21.6 years). More than half (54.1%) of the blood tests had at least one abnormal parameter, most often concerning the CBC. Anemia occurred in 5.3% of the population and was mostly normocytic. Microcytosis was mostly not associated with anemia (72.3% of cases). Lymphopenia occurred in 20.1% of the population and was mostly mild. Eosinophilia was present in 5.1% of the population and was never severe. Thrombocytopenia occurred in 0.7% of the population and was never severe. Serum levels of aminotransferases were elevated in 8.1% of the population. Fasting plasma glucose averaged 84 mg/dL (SD: 0.07) ranging from 64 to 123 mg/dL, was abnormal in 0.4% of the population, and one case of diabetes was diagnosed. Serum creatinine concentration was elevated in 0.7% of the population. Conclusion CBCs gave useful information but iron deficiency was common and insufficiently detected by this single analysis. Assessing aminotransferase levels without screening for viral hepatitis and systematic measurement of fasting plasma glucose levels did not appear to be efficient. In addition, the only interest in systematic measurement of creatinine serum levels was to obtain a reference level for long-term follow-up. In addition to the urinary dipstick, the systematic biological checkup at recruitment could be limited to a CBC with measurement of plasma ferritin levels and Hepatitis B virus serology, providing that any CBC abnormalities, in particular cytopenia, eosinophilia, and microcytosis, are systematically investigated. For a public health approach, systematic screening for other sexually transmitted infections could be proposed.


2020 ◽  
Author(s):  
Mei Zhang ◽  
Kun Yuan ◽  
Jingwen Liu ◽  
Haoran Zheng

Abstract Objective: Optical Coherence tomography (OCT) was employed to screen for maculopathy in the senile cataract population, investigate its incidence, and establish a healthy mode of management for maculopathy.Methods: A visual examination, slit-lamp microscope examination, direct ophthalmoscopic fundus examination, and a macular OCT examination were performed on 102 people with senile cataracts who were over 60 years in our hospital from January 1, 2019, to July 31, 2019. The demographic characteristics such as sex, age, physical examination mode: organization/individual, and routine physical examination items: presence or absence of hypertension, body mass index (BMI), blood biochemistry, total cholesterol, triglyceride (TG), fasting blood glucose and others.Results: Of the 102 subjects in the study, 28 were positive for maculopathy according to the OCT examination. Univariate analysis found that there were statistical differences between sexes and the presence of maculopathy (p<0.05). There were no significant differences in age, visual acuity, hypertension, BMI, fasting blood glucose, TG, total cholesterol, and cataract type (P>0.05). Multivariate logistic regression analysis, including all factors that might affect maculopathy, indicated that positivity for maculopathy and age (OR =2.549, 95%CI: 1.129–5.756, p<0.05) and gender (OR=3.907, 95%CI: 1.241–12.302, p<0.05) were related.Conclusion: The incidence of maculopathy in the senile cataract population was higher than that in the elderly population without cataract disease, and the proportion is 27.45%. Screening for maculopathy in the senile cataract population, especially among the elderly and females, should be improved.


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