Study the mRNA level of IL-27/IL-27R pathway molecules in kidney transplant rejection.

BACKGROUND: Renal transplantation stands as the sole remedy for individuals afflicted with end-stage renal diseases, and safeguarding them from transplant rejection represents a vital, life-preserving endeavor posttransplantation. In this context, the impact of cytokines, notably IL-27, assumes a critical role in managing immune responses aimed at countering rejection. Consequently, this investigation endeavors to explore the precise function of IL-27 and its associated cytokines in the context of kidney transplant rejection. METHODS: The study involved the acquisition of blood samples from a cohort of participants, consisting of 61 individuals who had undergone kidney transplantation (comprising 32 nonrejected patients and 29 rejected patients), and 33 healthy controls. The expression levels of specific genes were examined using SYBR Green Real-time PCR. Additionally, the evaluation encompassed the estimation of the ROC curve, the assessment of the relationship between certain blood factors, and the construction of protein-protein interaction networks for the genes under investigation. RESULTS: Significant statistical differences in gene expression levels were observed between the rejected group and healthy controls, encompassing all the genes examined, except for TLR3 and TLR4 genes. Moreover, the analysis of the Area Under the Curve (AUC) revealed that IL-27, IL-27R, TNF-α, and TLR4 exhibited greater significance in discriminating between the two patient groups. These findings highlight the potential importance of IL-27, IL-27R, TNF-α, and TLR4 as key factors for distinguishing between individuals in the rejected group and those in the healthy control group. CONCLUSIONS: In the context of kidney rejections occurring within the specific timeframe of 2 weeks to 2 months post-transplantation, it is crucial to emphasize the significance of cytokines mRNA level, including IL-27, IL-27R, TNF-α, and TLR4, in elucidating and discerning the diverse immune system responses. The comprehensive examination of these cytokines' mRNA level assumes considerable importance in understanding the intricate mechanisms underlying kidney rejection processes during this critical period.

Evaluation of morphology and angiogenesis of breast cancer in BALB/c mice using trypsin inhibitor from Cucumis melo seeds. In vitro and in vivo study.

Introduction: Melon seeds, as an excellent source of protease inhibitors, may have a protective role against tumor progression and angiogenesis. However, their effects on angiogenesis and the mechanism of their action against cancer progression remain elusive. This study aimed to investigate the effect of bioactive compounds of melon seed on the expression of angiogenesis genes in BALB/c mice with breast cancer. Material and methods: Trypsin inhibitor (TI) was purified from the seed powder of Cucumis melo. Half- maximal inhibitory concentration was determined for TI, extract of melon seed powder (EXT), and tamoxifen (TAM) by the (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test. Also, breast tumor was induced by subcutaneous injection of MC4-L2 cells in BALB/c inbred mice breast tissue. After tumor growth, mice were treated with TI, EXT, and TAM to examine their effects on the tumor characteristics and expression of angiogenesis-related genes including MMP-2, MMP-9, and vascular endothelial growth factor (VEGF) using the reverse transcription polymerase chain reaction method. Results: Trypsin inhibitor, EXT, TAM, and adjuvant treatment of TI + TAM resulted a reduction in expression of MMP-2, MMP-9, and VEGF. All treatments improved the breast tumor characteristics and the necrosis. The real-time polymerase chain reaction method verified the positive effects of the treatments on the breast cancer cell line and tumors. Conclusions: The results indicated that treatments with TI purified from Cucumis melo seeds and also combination therapy of TI and TAM can be considered as an alternative therapy in breast cancer patients. Further studies are warranted.

Effects of combined aerobic and anaerobic exercise training on cytokine profiles in patients with systemic lupus erythematosus (SLE); a randomized controlled trial.

BACKGROUND: Systemic lupus erythematosus (SLE) is an inflammatory rheumatic disease characterized by production of autoantibodies and organ damage. Elevated levels of cytokines have been reported in SLE patients. Physical activity could be considered one of the factors that affect the immune system status and function. The aim of the present study was to evaluate the effects of an 8-week supervised aerobic and anaerobic training program on the immune system of SLE patients through evaluation of serum cytokine levels. METHODS: This cross-sectional study included 24 SLE patients selected between September 2015 and March 2016. The patients were randomly divided into two groups, including exercise (n = 14) and control (n = 10) groups. The exercise group participated in an 8-week combined supervised exercise training program consisting of three times per week in 60-min exercise sessions. After collection of whole peripheral blood, peripheral blood mononuclear cells (PBMCs) were isolated from the blood samples. Following RNA extraction and cDNA synthesis, the expression levels of IFN-γ, TNF-α, IL-6, IL-2, IL-4, IL-5, IL-9, IL-10, IL-13, IL-17A, IL-17F, IL-21, and IL-22 were determined using in-house SYBER Green-based real-time polymerase chain reaction (PCR). Lastly, the data obtained were analyzed using t-test. RESULTS: The mean and standard deviation of age were 29.00 ± 3.19 and 21.50 ± 5.52 in the intervention and control groups, respectively. No significant differences were found among the mean serum levels of IFN-γ, IL6, IL-9, IL-17A, IL-17F and IL-21 among SLE patients in the intervention and control groups. The mean serum levels of TNF-α, IL2, IL-4, and IL-5 decreased significantly in the intervention as compared with the control group. The mean serum levels of IL-10, IL-13 and IL-22 significantly increased in the control group after eight weeks, as compared with the intervention group. CONCLUSIONS: Our findings indicated that the 8-week supervised aerobic and anaerobic training program could result in decreased inflammatory cytokines.

Comparison of FOXP3 and Interleukin 35 Expression Profiles in Kidney Transplant Recipients With Excellent Long-Term Graft Function and Acute Rejection.

OBJECTIVES: Transplant tolerance is defined as graft acceptance without long-term use of immunosuppressive agents. Regulatory T cells are involved in the maintenance of peripheral self-tolerance by actively suppressing the activation and expansion of autoreactive T cells. In the present study, we compared the expression profiles of forkhead box protein P3 (FOXP3) and interleukin 35 in kidney transplant recipients who had excellent long-term graft function under immunosuppression versus recipients who had acute rejection. MATERIALS AND METHODS: The 40 kidney transplant recipients included in this study were divided into 2 groups: 27 recipients with excellent long-term graft function and 13 recipients with acute rejection. After collection of whole peripheral blood, peripheral blood mononuclear cells were isolated from the blood samples. After RNAextraction and cDNAsynthesis from each collected sample, expression levels of interleukin 35 and FOXP3 were determined using in-house SYBER green-based real-time polymerase chain reaction. We used t tests to analyze data. RESULTS: Mean ages of recipients with excellent longterm graft function and recipients with acute rejection were 42.1 and 45.5 years, respectively. We found that FOXP3 and interleukin 35 expression levels were significantly increased in recipients with excellentlongterm graftfunction comparedwith recipientswith acute rejection. FOXP3 expression levels were significantly higher in those with excellent long-term graft function with graft survivalrate of <10 years,whereas interleukin 35 expression levels were significantly higher in patients with graft survival rate >10 years (P < .05). Expression levels of FOXP3 and interleukin 35 were greater in those from 35 to 50 years old versus with those in the other age ranges. CONCLUSIONS: Expression patterns of FOXP3 and interleukin 35 may have the potential to be used as prognostic biomarkers for kidney transplant outcomes.

Non-monotonic speed-dependence of microswimmers on wall distance.

While substrates naturally occur in most microswimmer experiments, their impact on the swimming performance is not well understood. In the present study, we functionalize substrates with polymer brushes of varying swelling properties, grafting densities and brush lengths to systematically modify and explore the substrate-swimmer interactions. Notably, the swimming speed does not monotonically change with brush thickness, but shows a distinct maximum at a certain intermediate thickness, which results from two counteracting factors: surface charge and surface roughness. The results show that the speed of thermophoretic microswimmers does not only depend on the particle properties but is also strongly influenced by the properties of the underlying substrate. This provides a route to control the speed of microswimmers via the underlying substrate, which could be applied in the future e.g. to design complex motility landscapes by patterning substrates with polymer brushes. It is expected that similar effects would occur for diffusio- and electrophoretic particles.

Circulating NKG2C + NK cell expressing CD107a/LAMP-1 subsets at the onset of CMV reactivation in seropositive kidney transplant recipients.

Cytomegalovirus (CMV) infection contributes to morbidity and mortality among kidney transplant recipients. Natural killer (NK) cells can battle against CMV in kidney transplant recipients (KTRs). This study aimed to analyze the association between CMV reactivation and the proportion of NK cell subsets and their activity. In a cross-sectional study, ten CMV reactivated KTRs, and ten non- CMV reactivated ones were recruited. Ten matched healthy controls were also included in this cohort. The presence of anti-CMV-IgG Ab in both KTR subgroups from seronegative donors and healthy controls was determined. The frequency of distinct subsets of memory-like NK cells was analyzed through NKG2C, NKG2A, and CD57 using flow cytometry. The activity of NK cells was evaluated after stimulation via coculture with K562 cell line and then assessment of the frequency of CD107a and granzyme B. The mRNA levels of transcription factors, including T-bet, EAT, and inflammatory proteins, including IFN-γ and perforin contributing to NK cell activation, were also evaluated. Results showed a significantly lower frequency of NKG2C + NKG2A-CD57+ NK cell population in CMV-reactivated KTRs compared to non-reactivated ones (P-value:0.003). NKG2C+ NK cells expressing CD107a/LAMP-1 significantly was increased in CMV-reactivated KTRs compared to non-reactivated ones (P-value: 0.0002). The mRNA level of IFN-γ had a significant increase in the CMV-reactivated KTRs vs. nonreactive ones (P-value: 0.004). Finally, evaluation of the NK cells' cytotoxicity and activity through assessment of CD107a/LAMP-1 expression and IFN-γ secretion may be helpful for the identification of the risk of CMV reactivation in KTRs.

Self-Propulsion of Janus Particles near a Brush-Functionalized Substrate.

Thermophoresis is a common mechanism that can drive autonomous motion of Janus particles under the right environment. Despite recent efforts to investigate the mechanism underlying the self-propulsion of thermophoretic particles, the interaction of particles with the substrate underneath the particle has remained unclear. In this work, we explore the impact of poly(N-isopropylacrylamide) (PNIPAM)-functionalized substrate with various chain lengths on the active motion of a single polystyrene particle half-coated with gold (Au-PS). We show how the modification of the substrate with polymer brushes enhances the particle velocity, where brush chain length plays a significant role as well. The results demonstrate the intrinsic dependence of particle velocity on the flow boundary condition and the thermo-osmotic slip at the interface.

Epidermal neural crest stem cell transplantation as a promising therapeutic strategy for ischemic stroke.

INTRODUCTION: Cell-based therapy is considered as promising strategy to cure stroke. However, employing appropriate type of stem cell to fulfill many therapeutic needs of cerebral ischemia is still challenging. In this regard, the current study was designed to elucidate therapeutic potential of epidermal neural crest stem cells (EPI-NCSCs) compared to bone marrow mesenchymal stem cells (BM-MSCs) in rat model of ischemic stroke. METHODS: Ischemic stroke was induced by middle cerebral artery occlusion (MCAO) for 45 minutes. Immediately after reperfusion, EPI-NCSCs or BM-MSCs were transplanted via intra-arterial or intravenous route. A test for neurological function was performed before ischemia and 1, 3, and 7 days after MCAO. Also, infarct volume ratio and relative expression of 15 selected target genes were evaluated 7 days after transplantation. RESULTS: EPI-NCSCs transplantation (both intra-arterial and intravenous) and BM-MSCs transplantation (only intra-arterial) tended to result in a better functional outcome, compared to the MCAO group; however, this difference was not statistically significant. The infarct volume ratio significantly decreased in NCSC-intra-arterial, NCSC-intravenous and MSC-intra-arterial groups compared to the control. EPI-NCSCs interventions led to higher expression levels of Bdnf, nestin, Sox10, doublecortin, ß-III tubulin, Gfap, and interleukin-6, whereas neurotrophin-3 and interleukin-10 were decreased. On the other hand, BM-MSCs therapy resulted in upregulation of Gdnf, ß-III tubulin, and Gfap and down-regulation of neurotrophin-3, interleukin-1, and interleukin-10. CONCLUSION: These findings highlight the therapeutic effects of EPI-NCSCs transplantation, probably through simultaneous induction of neuronal and glial formation, as well as Bdnf over-expression in a rat model of ischemic stroke.

MicroRNA Expression Profiles, Target Genes, and Pathways in Intervertebral Disk Degeneration: A Meta-Analysis of 3 Microarray Studies.

BACKGROUND: Determining the expression profile and target genes of microRNA (miRNA) would assist in determining the pathophysiologic pathways in intervertebral disk degeneration (IDD). The aim of this study was to determine the expression profile of miRNA in degenerated intervertebral disks compared with normal healthy intervertebral disks. METHODS: We conducted a meta-analysis of 3 available miRNA expression datasets to identify a panel of co-deregulated miRNA genes and overlapping biological processes in IDD. Degenerated intervertebral disks were compared with normal healthy disks. We selected 35 miRNA features common to all 3 platforms. Then, we calculated differential expression P values from our unpaired data using metaMA package in R statistical software according to the moderated t test method (Limma). Based on the P values (where the threshold was <0.05), a list of differentially expressed miRNAs was identified. RESULTS: After normalization and selection of common miRNA features across all 3 platforms, we found a total of 5 differentially expressed miRNAs, among which miR-574-3p, miR-199a-5p, and miR-483-5p were not identified in any individual studies. Our results revealed that miR-199a-5p, miR-574-3p, miR-551a, and miR-640 are commonly upregulated in IDDs compared with control disks, whereas miR-483 is commonly downregulated. Pathway analysis of identified dysregulated miRNAs indicated the involvement of extracellular matrix-receptor interaction, adherens junction, and transforming growth factor-beta signaling pathway in the pathogenesis of IDDs. Moreover, the network of predicted targets for these miRNAs identified most affected target genes as ERBB4 and CLTC. CONCLUSIONS: We found that the identified miRNAs through meta-analysis are candidate predictive markers for IDDs through different pathways.

Eugenol enhances proliferation and migration of mouse bone marrow-derived mesenchymal stem cells in vitro.

Mesenchymal stem cells (MSCs) have received considerable attention in regenerative medicine during the past decade. Eugenol is a natural and versatile vegetable molecule, which has a wide variety of therapeutic effects. Although different biological and pharmaceutical functions of Eugenol are well known, its effect on MSCs has not been studied yet. Therefore, this study was focused on investigating the effect of Eugenol on the proliferation and migration of bone marrow (BM)-derived MSCs in vitro. To do so, BM-MSCs were isolated from 4 to 8 weeks old NMRI mice. Cytotoxicity of Eugenol on MSCs was evaluated by MTT assay at 24, 48 and 72 h after treatment. In addition, its effect was assessed on the proliferation and migration of MSCs using wound healing assay in vitro and quantitative gene expression analysis for Oct4, Sox2, Cyclin-D1, Rex1, Tex10, Cxcr4, Vla4 and c-Met. Results showed that Eugenol reduced the number of MSCs in a dose- and time-dependent manner. The median inhibition concentration of Eugenol on MSCs was 400 µg/ml at 24 and 48 h and 200 µg/ml at 72 h after treatment. Moreover, about 90% viability of MSCs was detected at concentrations ≤12.5 µg/ml. The wound healing assay and gene expression analysis demonstrated that Eugenol promoted the migratory potential of MSCs through up-regulation of c-Met. Moreover, Eugenol has enhanced the proliferation of MSCs via over-expression of Sox2, Rex1 and Tex10. In conclusion, this study revealed that Eugenol enhances the proliferation and migration of MSCs, and thus this will be beneficial to the field of regenerative medicine.

Unveiling the Dynamics of Self-Assembled Layers of Thin Films of Poly(vinyl methyl ether) (PVME) by Nanosized Relaxation Spectroscopy.

A combination of nanosized dielectric relaxation (BDS) and thermal spectroscopy (SHS) was utilized to characterize the dynamics of thin films of poly(vinyl methyl ether) (PVME) (thicknesses: 7-160 nm). For the BDS measurements, a recently designed nanostructured electrode system is employed. A thin film is spin-coated on an ultraflat highly conductive silicon wafer serving as the bottom electrode. As top electrode, a highly conductive wafer with nonconducting nanostructured SiO2 nanospacers with heights of 35 or 70 nm is assembled on the bottom electrode. This procedure results in thin supported films with a free polymer/air interface. The BDS measurements show two relaxation processes, which are analyzed unambiguously for thicknesses smaller than 50 nm. The relaxation rates of both processes have different temperature dependencies. One process coincides in its position and temperature dependence with the glassy dynamics of bulk PVME and is ascribed to the dynamic glass transition of a bulk-like layer in the middle of the film. The relaxation rates were found to be thickness independent as confirmed by SHS. Unexpectedly, the relaxation rates of the second process obey an Arrhenius-like temperature dependence. This process was not observed by SHS and was related to the constrained fluctuations in a layer, which is irreversibly adsorbed at the substrate with a heterogeneous structure. Its molecular fluctuations undergo a confinement effect resulting in the localization of the segmental dynamics. To our knowledge, this is the first report on the molecular dynamics of an adsorbed layer in thin films.

Decoupling of Dynamic and Thermal Glass Transition in Thin Films of a PVME/PS Blend.

The discussions on the nanoconfinement effect on the glass transition and glassy dynamics phenomena have yielded many open questions. Here, the thickness dependence of the thermal glass transition temperature Tgtherm of thin films of a PVME/PS blend is investigated by ellipsometry. Its thickness dependence was compared to that of the dynamic glass transition (measured by specific heat spectroscopy) and the deduced Vogel temperature (T0). While Tgtherm and T0 showed a monotonous increase, with decreasing film thickness, the dynamic glass transition temperature (Tgdyn) measured at a finite frequency showed a nonmonotonous dependence that peaks at 30 nm. This was discussed by assuming different cooperativity length scales at these temperatures, which have different sensitivities to composition and thickness. This nonmonotonous thickness dependence of Tgdyn disappears for frequencies characteristic for T0. Further analysis of the fragility parameter showed a change in the glassy dynamics from strong to fragile, with decreasing film thickness.