Conversion of Unmodified Stem Cells to Pacemaker Cells by Overexpression of Key Developmental Genes.

Arrhythmias of the heart are currently treated by implanting electronic pacemakers and defibrillators. Unmodified adipose tissue-derived stem cells (ASCs) have the potential to differentiate into all three germ layers but have not yet been tested for the generation of pacemaker and Purkinje cells. We investigated if-based on overexpression of dominant conduction cell-specific genes in ASCs-biological pacemaker cells could be induced. Here we show that by overexpression of certain genes that are active during the natural development of the conduction system, the differentiation of ASCs to pacemaker and Purkinje-like cells is feasible. Our study revealed that the most effective procedure consisted of short-term upregulation of gene combinations SHOX2-TBX5-HCN2, and to a lesser extent SHOX2-TBX3-HCN2. Single-gene expression protocols were ineffective. Future clinical implantation of such pacemaker and Purkinje cells, derived from unmodified ASCs of the same patient, could open up new horizons for the treatment of arrythmias.

Maternal dietary diversity and nutritional adequacy in relation with anthropometric measurements of newborns at birth: a cohort study in Tehran city.

BACKGROUND: Maternal dietary intake during pregnancy plays an important role in fetal development and birth outcomes. The aim of the present study was to determine maternal dietary diversity and Nutritional adequacy in relation with anthropometric measurements of newborn at birth as a cohort study in Tehran city. METHODS: This prospective cohort study, was conducted by participation of 585 pregnant women referred to public health centers and hospitals covered by Shahid Beheshti, Tehran and Iran Universities in Tehran City. Using face-to-face interviews, general characteristics were obtained by questionnaire. Pre-pregnancy dietary intake was measured by a 168-item semi-quantitative food frequency questionnaire at the first visit, and dietary intake during pregnancy was measured by 2 non-consecutive 24-h food recall (one holiday and one regular day) at 31-34 weeks. Maternal height and weight were measured using standard tools and protocol at the first visit, and maternal weight at the end of pregnancy and data related to neonatal anthropometric indices were collected from mothers and neonates health records in the Sib electrical system. By applying SPSS software (version 23) the association was analyzed by linear regression with adjusting for confounding factors. P-value< 0.05 was considered as significant. RESULTS: Mean ± standard deviation of body mass index (BMI) of pre-pregnancy, pregnancy weight gain, BMI for age z-score (BAZ) at birth of infants were 24.52 ± 4.12, 12.16 ± 6.85 kg and - 0.61 ± 1.48, respectively. Mean ± SD of the Dietary Diversity Score (DDS) and Mean Adequacy Ratio (MAR) before and during pregnancy were 5.31 ± 1.11, vs.5.23 ± 1.42 and 289.85 ± 113.12 vs. 371.07 ± 197.28, respectively. After adjusting for confounding factors DDS in the third trimester of pregnancy was inversely correlated with WAZ (B = -0.16, 95% CI = - 0.23_0.30) and BAZ (B = - 0.24, 95% CI = - 0.06_0. 42) at birth, MAR of pre-pregnancy (B = - 0.001, 95% CI = - 0.002_0.00) and in the third trimester of pregnancy (B = - 0.18, 95% CI = - 0.35_0.004) were negatively associated with WAZ at birth. CONCLUSION: The findings showed that maternal nutritional status (dietary diversity and nutritional adequacy) before and during pregnancy were correlated with neonatal anthropometric indices at birth.

Sex Differences in Healthy Eating: Investigating the Moderating Effect of Self-Efficacy.

OBJECTIVES: This study aimed to evaluate sex differences in dietary habits and the moderating effect of self-efficacy on the adoption of a healthy diet. DESIGN: Cross-sectional study. SETTING: The healthy population of Tehran, Iran. PARTICIPANTS: A total of 262 participants from the general population with normal health status. MAIN OUTCOME MEASURE: The adoption of a healthy diet was based on the Healthy Eating Index-2015 (HEI-2015) scores and the moderating effect of self-efficacy on eating behaviors. ANALYSIS: Logistic regression analysis and multiple linear regression (moderation) analysis were conducted using PROCESS macro (version 3.5). RESULTS: The sex-based analysis revealed that females were more likely to adopt a healthy diet vs males (adjusted odds ratio, 1.85; 95% confidence interval, 1.02-3.35). The moderation analysis showed that eating behavior self-efficacy significantly moderated the relationship between sex differences and HEI-2015 scores (ΔR2 = 0.01; P = 0.033). Males with low self-efficacy scores had the highest difference in HEI-2015 with females with low self-efficacy scores, whereas the difference in HEI-2015 was very small in males and females with high self-efficacy scores. CONCLUSIONS AND IMPLICATIONS: Eating behavior self-efficacy had a significantly decreasing moderating effect on sex differences in the adoption of a healthy diet. Future research needs to clarify the impact of eating`` behavior self-efficacy in the adoption of a healthy diet, particularly in males, and to confirm the study's findings.

Unmodified autologous stem cells at point of care for chronic myocardial infarction.

BACKGROUND: Numerous studies investigated cell-based therapies for myocardial infarction (MI). The conflicting results of these studies have established the need for developing innovative approaches for applying cell-based therapy for MI. Experimental studies on animal models demonstrated the potential of fresh, uncultured, unmodified, autologous adipose-derived regenerative cells (UA-ADRCs) for treating acute MI. In contrast, studies on the treatment of chronic MI (CMI; > 4 wk post-MI) with UA-ADRCs have not been published so far. Among several methods for delivering cells to the myocardium, retrograde delivery into a temporarily blocked coronary vein has recently been demonstrated as an effective option. AIM: To test the hypothesis that in experimentally-induced chronic myocardial infarction (CMI; > 4 wk post-MI) in pigs, retrograde delivery of fresh, uncultured, unmodified, autologous adipose-derived regenerative cells (UA-ADRCs) into a temporarily blocked coronary vein improves cardiac function and structure. METHODS: The left anterior descending (LAD) coronary artery of pigs was blocked for 180 min at time point T0. Then, either 18 × 106 UA-ADRCs prepared at "point of care" or saline as control were retrogradely delivered via an over-the-wire balloon catheter placed in the temporarily blocked LAD vein 4 wk after T0 (T1). Effects of cells or saline were assessed by cardiac magnetic resonance (CMR) imaging, late gadolinium enhancement CMR imaging, and post mortem histologic analysis 10 wk after T0 (T2). RESULTS: Unlike the delivery of saline, delivery of UA-ADRCs demonstrated statistically significant improvements in cardiac function and structure at T2 compared to T1 (all values given as mean ± SE): Increased mean LVEF (UA-ADRCs group: 34.3% ± 2.9% at T1 vs 40.4 ± 2.6% at T2, P = 0.037; saline group: 37.8% ± 2.6% at T1 vs 36.2% ± 2.4% at T2, P > 0.999), increased mean cardiac output (UA-ADRCs group: 2.7 ± 0.2 L/min at T1 vs 3.8 ± 0.2 L/min at T2, P = 0.002; saline group: 3.4 ± 0.3 L/min at T1 vs 3.6 ± 0.3 L/min at T2, P = 0.798), increased mean mass of the left ventricle (UA-ADRCs group: 55.3 ± 5.0 g at T1 vs 71.3 ± 4.5 g at T2, P < 0.001; saline group: 63.2 ± 3.4 g at T1 vs 68.4 ± 4.0 g at T2, P = 0.321) and reduced mean relative amount of scar volume of the left ventricular wall (UA-ADRCs group: 20.9% ± 2.3% at T1 vs 16.6% ± 1.2% at T2, P = 0.042; saline group: 17.6% ± 1.4% at T1 vs 22.7% ± 1.8% at T2, P = 0.022). CONCLUSION: Retrograde cell delivery of UA-ADRCs in a porcine model for the study of CMI significantly improved myocardial function, increased myocardial mass and reduced the formation of scar tissue.

The effects of a multispecies probiotic supplement on inflammatory markers and episodic and chronic migraine characteristics: A randomized double-blind controlled trial.

BACKGROUND: The current study was designed to assess the effect of supplementation with a 14-strain probiotic mixture on episodic and chronic migraine characteristics. METHODS: Forty episodic and 39 chronic migraine patients who completed this randomized double-blind controlled trial received two capsules of multispecies probiotic or placebo. The migraine severity was assessed by visual analog scale (VAS). The number of abortive drugs consumed, migraine days, frequency and duration of attacks were recorded on paper-based headache diaries. Serum tumor necrosis factor alpha (TNF-α) and C- reactive protein (CRP) levels were measured at baseline and the end of the intervention. RESULTS: After a 10-week intervention, among episodic migraineurs the mean frequency of migraine attacks significantly reduced in the probiotic group compare to the placebo group (mean change: -2.64 vs. 0.06; respectively, p < 0.001). A significant reduction was also evident in the migraine severity (mean decrease: -2.14 in the probiotic group and 0.11 in the placebo group; p < 0.001). Episodic migraineurs who received the probiotic also showed significant reduction in abortive drug usage per week (mean change: -0.72; p < 0.001) compare to baseline, while there was no significant changes within the placebo group. In chronic migraine patients, after an 8-week intervention, the mean frequency of migraine attacks significantly reduced in the probiotic compared to the placebo group (mean change: -9.67 vs. -0.22; p ≤ 0.001). In contrast to the placebo, probiotic supplementation significantly decreased the severity (mean changes: -2.69; p ≤ 0.001), duration (mean changes: -0.59; p ≤ 0.034) of attacks and the number of abortive drugs taken per day (mean changes: -1.02; p < 0.001), in chronic migraine patients. We failed to detect any significant differences in the serum levels of inflammatory markers at the end of the study either in chronic or in episodic migraineurs. DISCUSSION: The results of this study showed that the 14-strain probiotic mixture could be an effective and beneficial supplement to improve migraine headache in both chronic and episodic migraineurs. Further research is required to confirm our observations.

A developmentally inspired combined mechanical and biochemical signaling approach on zonal lineage commitment of mesenchymal stem cells in articular cartilage regeneration.

Articular cartilage is organized into multiple zones including superficial, middle and calcified zones with distinct cellular and extracellular components to impart lubrication, compressive strength, and rigidity for load transmission to bone, respectively. During native cartilage tissue development, changes in biochemical, mechanical, and cellular factors direct the formation of stratified structure of articular cartilage. The objective of this work was to investigate the effect of combined gradients in cell density, matrix stiffness, and zone-specific growth factors on the zonal organization of articular cartilage. Human mesenchymal stem cells (hMSCs) were encapsulated in acrylate-functionalized lactide-chain-extended polyethylene glycol (SPELA) gels simulating cell density and stiffness of the superficial, middle and calcified zones. The cell-encapsulated gels were cultivated in a medium supplemented with growth factors specific to each zone and the expression of zone-specific markers was measured with incubation time. Encapsulation of 60 × 10(6) cells per mL hMSCs in a soft gel (80 kPa modulus) and cultivation with a combination of TGF-ß1 (3 ng mL(-1)) and BMP-7 (100 ng mL(-1)) led to the expression of markers for the superficial zone. Conversely, encapsulation of 15 × 10(6) cells per mL hMSCs in a stiff gel (320 MPa modulus) and cultivation with a combination of TGF-ß1 (30 ng mL(-1)) and hydroxyapatite (3%) led to the expression of markers for the calcified zone. Further, encapsulation of 20 × 10(6) cells per mL hMSCs in a gel with 2.1 MPa modulus and cultivation with a combination of TGF-ß1 (30 ng mL(-1)) and IGF-1 (100 ng mL(-1)) led to up-regulation of the middle zone markers. Results demonstrate that a developmental approach with gradients in cell density, matrix stiffness, and zone-specific growth factors can potentially regenerate zonal structure of the articular cartilage.

Experimental and computational investigation of the effect of hydrophobicity on aggregation and osteoinductive potential of BMP-2-derived peptide in a hydrogel matrix.

An attractive approach to reduce the undesired side effects of bone morphogenetic proteins (BMPs) in regenerative medicine is to use osteoinductive peptide sequences derived from BMPs. Although the structure and function of BMPs have been studied extensively, there is limited data on structure and activity of BMP-derived peptides immobilized in hydrogels. The objective of this work was to investigate the effect of concentration and hydrophobicity of the BMP-2 peptide, corresponding to residues 73-92 of the knuckle epitope of BMP-2 protein, on peptide aggregation and osteogenic differentiation of human mesenchymal stem cells encapsulated in a polyethylene glycol (PEG) hydrogel. The peptide hydrophobicity was varied by capping PEG chain ends with short lactide segments. The BMP-2 peptide with a positive index of hydrophobicity had a critical micelle concentration (CMC) and formed aggregates in aqueous solution. Based on simulation results, there was a slight increase in the concentration of free peptide in solution with 1000-fold increase in peptide concentration. The dose-osteogenic response curve of the BMP-2 peptide was in the 0.0005-0.005 mM range, and osteoinductive potential of the BMP-2 peptide was significantly less than that of BMP-2 protein even at 1000-fold higher concentrations, which was attributed to peptide aggregation. Further, the peptide or PEG-peptide aggregates had significantly higher interaction energy with the cell membrane compared with the free peptide, which led to a higher nonspecific interaction with the cell membrane and loss of osteoinductive potential. Conjugation of the BMP-2 peptide to PEG increased CMC and osteoinductive potential of the peptide whereas conjugation to lactide-capped PEG reduced CMC and osteoinductive potential of the peptide. Experimental and simulation results revealed that osteoinductive potential of the BMP-2 peptide is correlated with its CMC and the free peptide concentration in aqueous medium and not the total concentration.