The effect of Trimetazidine and Diazoxide on immunomodulatory activity of human embryonic stem cell-derived mesenchymal stem cell secretome

Comprehensive proteome profiling of the factors secreted by mesenchymal stem cells (MSCs), referred to as secretome, revealed that it consists of cytokines, chemokines, growth factors, extracellular matrix proteins, and components of regeneration, vascularization, and hematopoiesis pathways. Harnessing this MSC secretome for therapeutic applications requires the optimization of production of secretary molecules. A variety of preconditioning methods have been introduced, which subject cells to stimulatory molecules to create the preferred response and stimulate persistent effects. Pharmacological preconditioning uses small molecules and drugs to increase survival of MSCs after transplantation or prolong release of effective secretary factors such as cytokines that improve immune system responses. In this study, we investigated the effect of secretome of human embryonic-derived mesenchymal stem cells (hESC-MSCs) preconditioned with Trimetazidine (TMZ) and Diazoxide (DZ) on immunomodulatory efficiency of these cells in LPS-induced peripheral blood mononuclear cells (PBMCs). PBMCs were isolated from human peripheral blood and treated with concentrated hESC-MSC-derived conditioned medium and then, the secreted levels of IL-10, TNFα and IL-1β were assessed by ELISA after induction with LPS. The results showed that TMZ and DZ-conditioned medium significantly enhanced immunomodulatory potential of hESC-MSCs by increasing the secretion of IL-10, TNFα and IL-1β from LPS- induced PBMCs. We also found that hESC-MSCs did not secrete mentioned cytokines prior to or after the preconditioning with TMZ and DZ. In conclusion, our results implied that TMZ and DZ can be used to promote the immunomodulatory effects of hESC-MSC secretome. It is obvious that for applying of these findings in clinical demands, the potency of different pre-conditioned MSCs secretome on immune response needs to be more clarified.     

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Type 2 diabetes (T2D) is a condition with insufficient insulin production or in the setting of insulin resistance with many origins including intestinal microbiota-related molecular mechanism. Insulin-degrading enzyme (IDE) is responsible for insulin breakdown in various tissues and is known as a potential drug target for T2D. Here, we assessed the effects of cell-free supernatant (CFS) and UV-killed Lactobacillus casei IBRC_M10711 on IDE expression, IDE activity, and insulin degradation in Caco-2 cell line. It was found that CFS and UV-killed L. casei IBRC_M10711 led to lower expression of IDE. UV-killed L. casei IBRC_M10711 significantly inhibited IDE activity but CFS did not. Insulin degradation was affected with none of them. In conclusion, L. casei IBRC_M10711 is effective on IDE expression and its activity, but not on insulin degradation. Future studies are recommended to explore the effect of this probiotic on other substrates of IDE.     

Comparison of Fourier and wavelet transform procedures for examining the mechanomyographic and electromyographic frequency domain responses during fatiguing isokinetic muscle actions of the biceps brachii.

The primary purpose of the present study was to compare the fast Fourier transform (FFT) with the discrete wavelet transform (DWT) for determining the mechanomyographic (MMG) and electromyographic (EMG) center frequency [mean power frequency (mpf), median frequency (mdf), or wavelet center frequency (cf)] patterns during fatiguing isokinetic muscle actions of the biceps brachii. Seven men (mean+/-SD age=23+/-3 years) volunteered to perform 50 consecutive maximal, concentric isokinetic muscle actions of the dominant forearm flexors at a velocity of 180 degrees s(-1). Non-parametric "run" tests indicated significant (p<0.05) trends in the MMG and EMG signals for the 5th, 25th, and 45th muscle actions for all subjects, thereby confirming non-stationarity of the MMG and EMG signals. There were significant (p<0.05) correlations among the average normalized mpf, mdf, and cf values for contractions 1-50 for both MMG (r=0.671-0.935) and EMG (r=0.956-0.987). Polynomial regression analyses demonstrated quadratic decreases in normalized MMG mpf (R2=0.439), MMG mdf (R2=0.258), MMG cf (R2=0.359), EMG mpf (R2=0.952), EMG mdf (R2=0.914) and EMG cf (R2=0.888) across repetitions. The primary finding of this study was the similarity in the mpf, mdf, and cf patterns for both MMG and EMG, which suggested that, despite the concerns over non-stationarity, Fourier based methods are acceptable for determining the patterns for normalized MMG and EMG center frequency during fatiguing dynamic muscle actions at moderate velocities.     

Mechanomyographic and electromyographic responses of the vastus medialis muscle during isometric and concentric muscle actions

The purpose of this study was to examine the patterns for the mechanomyographic (MMG) and electromyographic (EMG) amplitude and mean power frequency (MPF) vs. torque relationships during submaximal to maximal isometric and isokinetic muscle actions. Seven men (mean +/- SD age, 22.4 +/- 1.3 years) volunteered to perform isometric and concentric isokinetic leg extension muscle actions at 20, 40, 60, 80, and 100% of maximal voluntary contraction (MVC) and peak torque (PT) on a Cybex II dynamometer. A piezoelectric MMG recording sensor was placed between bipolar surface EMG electrodes on the vastus medialis. Polynomial regression and separate 1-way repeated-measures analysis of variance were used to analyze the EMG amplitude, MMG amplitude, EMG MPF, and MMG MPF data for the isometric and isokinetic muscle actions. For the isometric muscle actions, EMG amplitude (R(2) = 0.999) and MMG MPF (R(2) = 0.946) increased to MVC, mean MMG amplitude increased to 60% MVC and then plateaued, and mean EMG MPF did not change (p > 0.05) across torque levels. For the isokinetic muscle actions, EMG amplitude (R(2) = 0.988) and MMG amplitude (R(2) = 0.933) increased to PT, but there were no significant mean changes with torque for EMG MPF or MMG MPF. The different torque-related responses for EMG and MMG amplitude and MPF may reflect differences in the motor control strategies that modulate torque production for isometric vs. dynamic muscle actions. These results support the findings of others and suggest that isometric torque production was modulated by a combination of recruitment and firing rate, whereas dynamic torque production was modulated primarily through recruitment.     

Gender comparisons of mechanomyographic amplitude and mean power frequency versus isometric torque relationships

This study compared the patterns of mechanomyographic (MMG) amplitude and mean power frequency vs. torque relationships in men and women during isometric muscle actions of the biceps brachii. Seven men (mean age 23.9 +/- 3.5 yrs) and 8 women (mean 21.0 +/- 1.3 yrs) performed submaximal to maximal isometric muscle actions of the dominant forearm flexors. Following determination of the isometric maximum voluntary contraction (MVC), they randomly performed submaximal step muscle actions in 10% increments from 10% to 90% MVC. Polynomial regression analyses indicated that the MMG amplitude vs. isometric torque relationship for the men was best fit with a cubic model (R(2) = 0.983),,where MMG amplitude increased slightly from 10% to 20% MVC, increased rapidly from 20% to 80% MVC, and plateaued from 80% to 100% MVC. For the women, MMG amplitude increased linearly (r(2) = 0.949) from 10% to 100% MVC. Linear models also provided the best fit for the MMG mean power frequency vs. isometric torque relationship in both the men (r(2) = 0.813) and women (r(2) = 0.578). The results demonstrated gender differences in the MMG amplitude vs. isometric torque relationship, but similar torque-related patterns for MMG mean power frequency. These findings suggested that the plateau in MMG amplitude at high levels of isometric torque production for the biceps brachii in the men, but not the women, may have been due to greater isometric torque, muscle stiffness, and/or intramuscular fluid pressure in the men, rather than to differences in motor unit activation strategies for modulating isometric torque production.     

Occurrence and physico-serological properties of potato virus S (PVS) in Iran

1818 collected samples of potato plant showing virus infection symptoms from 85 fields were tested for PVS infection using DAS-ELISA. Average of infection to this virus varied from 0 to 100%. Least infection was belonging to fields with new introduced varieties. On the other hand native and old introduced cultivars showed heavy infection. In field condition, PVS infected plants didn't show very obvious symptoms, so some infected plants may be missed in field sample collecting. The physical properties of 3 isolates, Avaj, Stanboly and Agria No 15 were determined. TIP 55-60 degrees C, DEP 10(-3) and Liv measured 3-4 days. Ouchterlony agar double diffusion test using SDS was useful for virus detection and precipitation lines didn't show any spur between isolates, although isolates differs slightly in symptomatology. SDS-Page and Western blotting methods used successfully for virus detection and determining and measuring viral protein components.     

Chitinase gene transformation through Agrobacteriumand its explanation in soybean in order to induce resistance to root rot caused by Rhizoctonia solani

Chitinase gene (chi) of bean which has been cloned in recombinant binary plasmid vector, pBI121 with 35s promoter of Cauliflower mosaic virus (CaMV), were used for transformation of soybean using strain LBA4404 of Agrobacterium. The plasmid contained nptII gene that is a resistant gene to kanomycin as selector marker and Gus gene as reporter. Cotyledon explants of Williams and Clark cultivars were inoculated by Agrobacterium suspension with pBI121 and were cultured in regeneration medium. After complete regeneration of explants to seedling in B5 medium amended with kanomycin, polymerase chain reaction analysis were conducted to ensure conjugation of nptII, Gus, CHN genes in transformants seedling of soybean. Results showed that some lines of soybean contained Gus and CHN genes. More ever, chitinase activity in leaf extract of transformed soybean lines was significantly more than untransformed soybean, exception one sample. Bioassay of chitinase activity of transgenic lines on in vitro condition prevented mycelial growth of Rhizoctonia solani in comparison with untransformed control leaf extract.     

Synthesis and characterization of dual function vanadyl, gallium and indium curcumin complexes for medicinal applications

Novel bis[4-hydroxy-3-methoxyphenyl]-1,6-heptadiene-3,5-dione (curcumin) complexes with the formula, ML(3), where M is Ga(III) or In(III), or of the formula, ML(2) where M is [VO](2+), have been synthesized and characterized by mass spectrometry, infrared and absorption spectroscopies, and elemental analysis. A new ligand, bis[4-acetyl-3-hydroxyphenyl]-1,6-heptadiene-3,5-dione (diacetylbisdemethoxycurcumin, DABC) was similarly characterized; an X-ray structure analysis was performed. Vanadyl complexes tested in an acute i.p. testing protocol in STZ-diabetic rats showed a lack of insulin enhancing potential. Vanadyl complexes were, however, more cytotoxic than were the ligands alone in standard MTT (3-[4,5-dimethylthiazole-2-yl]ate, -2,5-diphenyl-tetrazolium bromide) cytotoxicity testing, using mouse lymphoma cells. With the exception of DABC, that was not different from VO(DABC)(2), the complexes were not significantly different from one another, with IC(50) values in the 5-10 microM range. Gallium and indium curcumin complexes had IC(50) values in the same 5-10 microM range; whereas Ga(DAC)(3) and In(DAC)(3) (where DAC=diacetylcurcumin) were much less cytotoxic (IC(50)=20-30 microM). Antioxidant capacity was decreased in VO(DAC)(2), Ga(DAC)(3), and In(DAC)(3), compared to vanadyl, gallium and indium curcumin, corroborating the importance of curcumin's free phenolic OH groups for scavenging oxidants, and correlated with reduced cytotoxic potential.     

Propranolol ameliorates and epinephrine exacerbates progression of acute and chronic viral myocarditis

Recent studies point to important interactions between proinflammatory cytokines and neurohumoral mediators in heart failure. Here we investigate the influence of the beta-adrenergic system on cytokines and neurohumoral factors and the sequelae of viral myocarditis. In an experimental model with virus-infected BALB/c mice, we studied the acute and chronic effects of epinephrine and propranolol on myocardial morphology, cytokine gene expression, and survival. BALB/c mice were inoculated with the encephalomyocarditis virus (EMCV) or sham inoculated with saline and followed for 30 days. Epinephrine increased the severity of inflammatory cell infiltration and myocardial necrosis induced by EMCV. Gene expression of TNF-alpha, IL-6, and IL-10 was markedly enhanced by epinephrine in EMCV-inoculated mice. Survival rate after 30 days was reduced to 40% in epinephrine-treated EMCV-inoculated mice compared with 70% in untreated EMCV-inoculated mice (P < 0.05). Treatment with the beta-blocker propranolol significantly decreased mortality, myocardial necrosis, and infiltration of inflammatory cells in EMCV-inoculated mice. Propranolol also suppressed gene expression of TNF-alpha, IL-6, and IL-10. A single dose of epinephrine 120 days after EMCV inoculation caused sudden death in 70% of infected mice; propranolol significantly reduced incidence of death to 33%. These results indicate that acute and chronic stages of viral myocarditis are modulated by the beta-adrenergic system and its interactions with proinflammatory cytokines.