Background and Aims

To establish a method for isolation and induction of mouse bone marrow-derived macrophages (BMDM) and to evaluate the effect of high glucose on macrophage polarization.

Methods

BMDMs were isolated for the femur and tibia of 6-8 weeks C57BL/6 mice, and identified by flow cytometry, then be divided into M1 (induced by LPS and IFN-γ), M2(induced by IL-4), normal glusose (5.6 mmol/l), high glucose (30 mmol/l) and osmotic pressure control group (normal glucose+mannitol 24.4mmol/L), stimulated by high glucose for 24 and 48 hours, collect cells for flow cytometry and qPCR detection of M1 and M2 cell surface marker expression.

Results

BMDMs were isolated successfully, the proportion of CD11b and F4/80 double positive cells was above 95%; After induction, In M1 subgroup, the flow cytometry results suggested that CD86 positive rate increased significantly, qPCR results showed that IL-1b, iNOS expression increased significantly; in M2 subgroup, CD206 positive rate increased significantly, the expression of Arg1 and Ym1 were significantly increased. After stimulation with high glucose, with the prolongation of high glucose stimulation time, the ratio of CD86 positive rate in BMDMs increased gradually, while the positive rate of CD206 did not increase significantly; qPCR results showed that with high glucose stimulation over time, M1 macrophage labeled IL-1b, iNOS expression was significantly increased (p <0.05).

Conclusions

We have successfully established a method for isolation and induction culture of BMDMs. After 48 hours of high glucose stimulation, macrophages will be polarized to M1.

Background and Aims

Abdominal obesity strongly associates with multiple metabolic abnormalities (dyslipidemia, insulin resistance [IR], increased fasting glucose and impaired glucose tolerance) and higher cardiovascular (CV) risk. The aim of the study: to investigate the number of circulating EPCs in patients with asymptomatic cardiac dysfunction and different phenotypes of obesity.

Methods

The study was retrospectively evolved 46 patients with asymptomatic chromic heart failure (left ventricular ejection fraction 40%-49%) and established abdominal obesity (47 patients with metabolically unhealthy obesity [Met-UHO] and 42 subjects with metabolically healthy obesity [Met-HO]) from the large cohort of dismetabolic patients (n=268). High-Definition Fluorescence Activated Cell Sorter methodology was performed for measurement of the number of circulating endothelial progenitor cells co-expressed CD45, CD34, CD14, CD309, and Tie-2 antigens.

Results

A significant difference between number of circulating progenitor cells labeled CD45-CD34+ and CD14+CD309+ in Met-UHO and Met-HO patients was found. In contrast, Met-UHO patients had a significantly lower level of circulating CD14⁺ Tie-2⁺ cells and СD309⁺ Tie-2⁺cells compared with Met-HO individuals. In multivariate logistic regression analysis we found that HOMA-IR, hs-CRP, and number of CV risk factorswere independent predictors for depletion in numerous of circulating progenitor cells with immune phenotypes CD309/Tie2+ cells and CD14/Tie2+.

Conclusions

We found that the lowered circulating number of CD309/Tie2+ cells / CD14/Tie2+ cells produces the well balanced discrimination on Met-UHO development in Met-HO patients with co-existing preserved left ventricular ejection fraction than other models based on conventional CV risk factors.

Background and Aims

TCF7L2 gene encodes a transcription factor expressed in pancreatic β cells that regulates insulin production and processing. It is assumed that type 2 diabetes risk gene TCF7L2 affects the response to diet therapy.

We aimed to assess an effect of the single nucleotide polymorphism (SNP) rs7903146 T/C in the TCF7L2 gene on the change of bioimpedance parameters in overweight patients who followed a generally accepted for obesity therapeutic diet for 3 months.

Methods

The study implicated 17 overweight or obese patients (15 women and 2 men) aged 23 to 60 years (average BMI on admission – 34.1±5.6 kg/m²). Within 3 months, all patients have followed a balanced therapeutic diet with the exception of easily digestible and limited digestible carbohydrates and fats. Patient genotyping data were compared with the European population (Project "1000 Genomes", n = 503). DNA was extracted from a venous blood. Gene polymorphisms were identified by real-time PCR (CFX96, USA).

Results

The observed genotype distribution was consistent with the Hardy-Weinberg equilibrium (p>0.05) and was not significantly different from the European frequency distribution. CC homozygotes of the SNP demonstrated a significant decrease in body cell mass (BCM) and total water (in kg) after 3 months of diet therapy compared to the T-allele carriers (p=0.013 and p=0.018, respectively).

Conclusions

Carriers of risk allele T demonstrated a better response to diet therapy manifested in BCM gain. More detailed study may suggest a suitable diet to compensate the adverse effect of genotype in risk allele carriers.

Background and Aims

Background and Aims

The combination of peptide YY (PYY) and glucagon-like peptide-1 (GLP-1) has been proposed as a potential treatment for diabetes and obesity. Developing such co-formulation for a commercial product is very challenging due to different formulation conditions required to maintain the two peptides in stable form. The aim of the study reported here was to develop a stable co-formulation of PYY and liraglutide, a selected GLP-1.

Methods

Co-formulations were prepared in glass vials and the stability was assessed by reversed-phase chromatography (RP-HPLC) methods for the respective peptides and by a visual assessment compliant with the 2.9.20 EP Monograph. Stability was tested following storage at 30ºC.

Results

The initial screening demonstrated the incompatibility of PYY and liraglutide, and stability of the co-formulation in the composition of the marketed liraglutide product was found to be poor. Screening of various excipients and stabilising conditions resulted in identification of pharmaceutically acceptable compositions that enabled a stable liquid co-formulation of PYY and liraglutide

Fig. 1. Stability of PYY (left) and liraglutide (right) at 30ºC accelerated study in the Control formulation and a stabilising formulation developed in this study. The Control formulation failed the visual assessment at 4 weeks and 8 weeks; the stabilised formulation passed the visual assessment at all time-points.

Conclusions

The present study demonstrated the feasibility of developing a stable co-formulation of PYY and liraglutide. The stabilising compositions were based on excipients with history of use in injectable products for human use, which significantly de-risks further development plans.

Background and Aims

DiaGone™ is an endoscopic device which utilizes precisely controlled laser technology to target the duodenal submucosal neural plexi with the aim of improving glucose control by modulating the gastrointestinal neurohumoral axis.

Methods

Nine subjects (5 males) with obesity (BMI 34.0±4.6 kg/m²) and type 2 diabetes mellitus (T2DM) insufficiently controlled on metformin were included in this first-in-human trial in order to assess the efficacy and safety of DiaGone™. Biochemical and anthropometric parameters were assessed at baseline and 3 and 6 months after the procedure and a standard liquid meal test was performed at baseline and 3 months after the procedure.

Results

DiaGone™ significantly decreased fasting glucose (12.4±3.5 vs. 9.5±2.0 vs. 9.7±2.7 mmol/l for baseline vs. 3 vs. 6 months, p<0.01) and HbA_1C (78.3±13.3 vs. 64.9±6.4 vs. 64.8±7.1 mmol/mol, p<0.01) as well as. Glucose AUC during the 150 min meal test 3 months after the procedure showed a reduction of 20% from baseline (43,435 vs. 34,096, p<0.01). No significant changes were observed in AUC for insulin. No adverse events related to the procedure or significant changes in weight were reported throughout the 6-month period.

Conclusions

These pilot results suggest that endoscopic duodenal submucosal laser ablation using the DiaGone™ device is associated with improvements in both baseline and postprandial glycemia as well as HbA_1C with no significant changes in insulin levels or body weight while having a favorable safety and tolerability profile.

Supported by MHCZ-DRO („Institute for Clinical and Experimental Medicine – IKEM, IN 00023001“) and RVO VFN64165.

Background and Aims

Obesity represents health, social and economic threat for most of countries worldwide. Bariatric surgery of obesity is the most effective long-term treatment modality for patients suffering from higher degrees of obesity.

The aim was to use new technologies for treating obesity and preventing diabetes: gastric balloon and laparoscopic bandage of gastric.

Methods

We examined 508 obesity patients (379 females and 129 males) aged 19.9 to 78.5 years. The mean age was 48.5±29.5 years. BMI varied from 30.2 to 60.1 kg/m². In 68 patients with BMI 30.0 - 40.0 kg / m² was used gastric balloon for 6 months. We used laparoscopic bandage of gastric (Obtech Medical AG) in 54 patients with BMI more than 40.0 kg/m².

Results

Obesity-related comorbidities were revealed in 92.3% of patients: arterial hypertension – 87.6%; back pain – 79.9%; type 2 diabetes – 31.1%; osteoarthritis – 26.9%; reflux esophagitis – 24.5%; dysmenorrhea – 23.4%; sleep apnoe – 19.1%. Weight loss after use of gastric balloon was from 23 to 38 kg (mean 30.5±15.2 kg), max after 6 months. Complications were revealed in 6 (8.8%) patients. Weight loss in patients after laparoscopic bandage of gastric was from 23 to 58 kg (mean 40.5±16.7 kg). After 1 year BMI was normal in 32.7% this patients. Complications were revealed in 4 patients. Glycaemia, cholesterol, triglycerides significantly decreased in two groups.

Conclusions

New technologies for treating obesity, gastric balloon and laparoscopic bandage of gastric, have positive effects on weight loss, preventing diabetes and improving of obesity-related comorbidities.

Background and Aims

Aims: The association of the endothelial dysfunction(ED) marker - Plasminogen Activator Inhibitor-1(PAI-1), insulin resistance(IR) indicators and the cardiovascular risk(CVR) to study in patients with prediabetes and without cardiovascular event(CVE).

Methods

Materials and methods. A case-control study conducted, including 406 people, aged 18 to 65 years. The glycated hemoglobin(HbA1c,%), C-peptide, insulin, biomarker PAI-1, IR index HOMA measured. After clinical examination, patients were divided into 3 groups: 1 – the control group n=20, 2 group – the prediabetes without CVE n=80, 3 group - the prediabetes with CVE n=30.

Results

Results. Significant differences the PAI-1 level were found between the 2 group(PAI-1; Me -29754,3 pg/ml) and the 3 group(Me-36867,2 pg/ml) in comparison with the control group(Me-24238,5 pg/ml; p=0,000).We considered Means±SD the PAI-1 level based on SCORE scale in the 2 group: PAI-1in the low CVR group SCORE(<1points) was 32345,1±24191,7 pg/ml; in the high CVR group SCORE(>1points) Mean was-38570,7±22768,7 pg/ml;p=0,000. The relationship between PAI1 and SCORE points found(r=0,23; р=0,001).The relationship PAI-1 with glucagon(r=0,23; р=0,001), HOMA index(r=0,27; р=0,000), С-peptide(r=0,25; р=0,001), fasting glucose(r=0,23; р=0,000) found. The relationship SCORE points with insulin(r=0,43; р=0,001), HOMA index(r=0,47; р=0,000), С-peptide(r=0,38; р=0,001), LDL(r=0,4; р=0,000), triglycerides(r=0,32; р=0,000) determined. On increasing PAI-1 biomarker level affected elevation SCORE points on 2,6 fold(р=0,05), HbA1c on 1,3 fold(р=0,05), hyperglucemia on 1,2 fold(р=0,02) based on results polynomial regression.

Conclusions

Conclusions. We found the association of the PAI-1 biomarker, IR indicators and the CVR in patients with prediabetes. Indicating that with an increase of the PAI-1, IR increases and thereby increases the CVR and ED in these patients.

Background and Aims

ANGPTL8 has been reported to be a regulator of lipid metabolism, and it is associated with insulin resistance (IR) and metabolic syndrome (MetS). We investigated whether ANGPTL8 plays a role in MetS.

Methods

ANGPTL8 and adiponectin concentrations were measured in PCOS patients with or without MetS and in their corresponding healthy controls. The association of circulating ANGPTL8 with adiponectin and other parameters was also examined.

Results

Circulating ANGPTL8 concentrations were higher in PCOS women with MetS than in those without MetS and in the controls (P <0.01). ANGPTL8 was positively correlated with age, BMI, FAT%, WHR, SBP, TG, FBG, HbA1c, Fins, and HOMA-IR (all P <0.01) in the study populations and negatively associated with adiponectin and M-values (P <0.001). In addition, ANGPTL8 was positively correlated with PRL, LH, TEST and FAI and negatively correlated with SHBG (all P <0.01). ROC curve analyses showed that the AUC_MetS was 0.87 (P < 0.001), with a sensitivity of 92.4% and specificity of 75.4%, and the AUC_IR was 0.82 (P < 0.01), with a sensitivity of 76.4% and specificity of 75.6%.

Conclusions

ANGPTL8 levels progressively decrease from PCOS patients with MetS to those without MetS and may be a serum marker associated with the degree of metabolic disorders.

Background and Aims

Even though diabetes patients exhibit an increase in oxidative stress, its correlation with declining renal function is not fully understood. The purpose of this study was to determine whether hydrogen peroxide, 8-isoPGF2α, and oxLDL-β2GP1 complexes correlated with renal function decline as shown by eGFR in type 2 diabetes mellitus (T2DM) patients.

Methods

The design of this study was cross-sectional. A total of 227 T2DM patients (134 normoalbuminuria patients and 93 albuminuria patients) were divided based on the eGFR value. Group 1 was patients with eGFR > 60 mL/min/1.73 m² (n=195) and group 2 was patients with eGFR ≤ 60 mL/min/1.73 m² (n=32). T2DM patients’ serum and urine specimens were analyzed to measure their serum creatinine and eGFR based on CKD-EPI equation. Serum oxLDL-β2GPI complexes and urine 8-isoPGF2a were measured by ELISA and urine hydrogen peroxide was measured by Ferrous Ion Oxidation Xylenol Orange 1 (FOX-1).

Results

The results showed that oxLDL/β2GPI was higher in group 2 (0.60±0.08 units/mL) than group 1 (0.59±0.04 units/mL) significantly (p=0.04). In contrary, hydrogen peroxide and 8-iso-PGF2α were lower in group 2 than group 1), but not statistically significant. However, no significant correlation between eGFR with oxLDL/β2GPI and the other two parameters. Only oxLDL/β2GPI and hydrogen peroxide showed a significant correlation (r=0.145, p=0.029).

Conclusions

In conclusion, serum oxLDL-β2GP1 has a role in the kidney function decline in T2DM patients that contribute to nephropathy diabetes and showed correlation with hydrogen peroxide, but not for 8-iso-PGF2α.

Background and Aims

The aim of our study is to assess the efficacy of lifestyle modification and picolinate chromium intake in preventing diabetes mellitus type 2 (DM 2)

Methods

The study included 100 patients (69 m 258 f) 25-65 years with impaired glucose tolerance (IGT) and newly diagnosed DM 2. Patients were divided into 2 groups matched by age, weight, body mass index (BMI), waist-to-hip ratio (WHR). Research group included 54 patients who carried out recommendations of a balanced diet, physical activity and picolinate chromium was taken 200mkg per day for 1 month. Control group included 46 patients who did not lifestyle modification. The study was 24 weeks. We measured fasting plasma glycated hemoglobin (HbA1c,%) and related to fasting leptin (FL).

Results

: Patients of the research group demonstrated reduction of BMI (-3.2±2.1 kg/m²) and WHR (-0.02±0.025) (p<0.01 for all) and positive dynamics of HbA1c ( p<0.001). Persons of the control group had increase BMI, WHR as also HbA1c elevation (p<0.05). The main novel finding was that median serum leptin in research group decreased on -23.9% (p<0.01) and increased in control group on +27.6% (p<0.01). Among subjects with IGT from the research group, HbA1c normalized in 49.3% (p<0.001) and serum leptin levels decreased on 26.9% (p<0.01). In control group HbA1c normalized in 4.5% (p<0.01). Among patients of the research group was a reduction of DM 2 by 11.9% and an increase in the control group by 35.1%.

Conclusions

Lifestyle modifications with picolinate chromium intake leads to the restoration of glucose homeostasis and preventing DM 2

Background and Aims

Nowadays a huge number of genetic polymorphisms, associated with insulin resistance have been identified. However, their prevalence may have regional differences. Our aim was to study the possible association of the FTO rs9939609, PPARG rs180128, PPARGC1A rs8192678 polymorphisms with the development of type 2 diabetes mellitus (T2DM) and early carbohydrate metabolism disorders in the residents of the Republic of Tatarstan.

Methods

The study included 272 obese patients: a group of patients with T2DM and a group with prediabetes. The control group was taken from the 1000 genomes database (European population, n=503). DNA was isolated from leukocytes, followed by the determination of alleles and genotypes using real-time PCR.

Results

Bearing of the AA genotype and A allele of the FTO gene polymorphism increases the risk of both T2DM (OR=1,73, p=0,0006; OR=1,73, p<0,05, respectively) and prediabetes (OR=2,93, p<0,05; OR=2,51, p<0,05, respectively) development. Bearing of the G allele and GG genotype of the PPARG gene polymorphism significantly increases the risk of T2DM (OR=1,63, p=0,009; OR=7,29, p=0,0002, respectively), but no association with prediabetes was identified. PPARGC1A gene polymorphism showed a significant effect of the C allele and CC genotype both in T2DM (OR=1,58, p=0,003; OR=1,86, p=0,007, respectively) and prediabetes (OR=1,63, p=0,001; OR=1,87, p=0,005, respectively).

Conclusions

The associations of aforesaid polymorphic markers with the T2DM development in the Republic of Tatarstan have been proved, as well as the associations of the FTO and PPARGC1A genes polymorphisms with the development of early carbohydrate metabolism disorders.

Background and Aims

was to estimate the affect of complex treatment with kinesiotherapy on body weight loss and muscle function in patients with obesity.

Methods

80 men and women aged 21-69 years old with obesity were enrolled in the study. The complex kinesiotherapy included interactive sensorimotor trainings. Weight, WC, HC, fall number were measured at baseline and after the treatment was completed. Muscle strength and walking speed functional tests results assessment were performed.

Results

There was a significant reduction in body weight (111.3±24.4 kg at baseline vs 107.9±23.1 kg in 3 weeks; р=0,000), in BMI (40.3±8.1 vs 39.1±7.7 kg/m²; р=0.000),i n treated obese patients. 10-meters-walk speed increased from 0.84±0.15 m/sec at baseline to 0.88±0.17 m/sec to (р=0.000). Up-and-go test improved from 8.4±2.1 to 7.9±2.09 sec (р=0.000). We registered endurance to static loading in abdomen muscles from 13.1±9.7 to 16.49±12.8 sec (р=0.000) and in back muscles from 14.8±11.9 sec to 18.6±14.9 sec (р=0.000). The endurance to dynamic loading increased in abdomen muscles from 29.9±11.2 to 34.84±11.93 times (р=0.000) and also in back muscles from 9.1±7.4 to 12.2±9.2 times (р=0.000). Fall namber markably decreased from 0.14 ±0.34 at baseline to 0.0 (95%CI: 0.02; 0.25) after treatment.

Conclusions

Investigated complex treatment with kinesiotherapy methods promotes body weight loss, WC and HC reduction in obesity. Special training of obese patients is associated with increasing in gate speed and lower extremities muscle strength, and it also causes improvement in static and dynamic loading endurance of back and abdomen muscles. Those changes may probably improve balance function and decrease risk of falling in obese patients.

Background and Aims

Prevalence of type 2 Diabetes in Mexico is high. Early detection and intervention of the condition, particularly in children and adolescents will have a great impact in the wellbeing of individuals and reduce the cost of treatment. CARDIGAN aims to test whether Criticality Analysis (CA) can be applied to gait data to give a reliable and cost-effective way to detect individuals at risk of developing type 2 diabetes.

Methods

Gait data and clinical data of overweight children was collected using portable Inertial Measurement Units in a 6-week trial, conducted by Hospital Infantil de Mexico Federico Gomez. Data analysis is now being performed in a semi-blind manner using a novel machine learning approach based on CA.

Results

The results obtained from the CA show visible differences in the gait patterns among the obese participants, and even more pronounced differences are seen compared with the control group. Data from each week will be analysed to track the progression of the participants. All of these results will be cross-referenced with the clinical data obtained.

Conclusions

The use of Criticality Analysis of gait as a means of diabetes assessment is evaluated in order to track the progression of the participants’ condition, and detect if they are at risk of developing type 2 diabetes. This will aid in determining the appropriate course of intervention.

This work was supported by an Institutional Links grant, 432368181. The grant is funded by the UK Department for Business, Energy and Industrial Strategy and delivered by the British Council. For further information, please visit www.newtonfund.ac.uk.