Magnetic Field Exposure Modulates the Anti-Inflammatory Efficiency of Minocycline in Rats with Peripheral Acute Inflammation.

BACKGROUND AND OBJECTIVE: Microglial activation in spinal cord is key contributor and its inhibition by Minocycline (MCN) can result in anti-inflammatory actions. Effect of pulsed magnetic field (PMF) in living system is a very complex process and many biological and cellular processes can play key roles. In this study aimed to reveal the roles of PMF exposure on anti-inflammatory potentials of MCN treatment by evaluating the inflammatory profiles of either inflamed site or spinal cord. METHODS: In this study, we investigated the anti-inflammatory effects of PMF, MCN or their combination treatments in rats with carrageenan (CG)-induced peripheral inflammation by examining the cardinal signs, hyperalgesia, allodynia, edema and fever. The levels of various inflammation markers (tumor necrosis factor-α), interleukin (IL)-1ß, IL-6, IL-17, IL-4, IL-10, C-C motif chemokine ligand3 (CCL3), C-X-C motif chemokine ligand1 and myeloperoxidase were also measured in paw and spinal cord tissues. RESULTS: CG induced inflammation caused edema, fever, and hypersensitivities. MNC or PMF treatments ameliorated these responses by suppressing pro-inflammatory markers in both inflamed paw and spinal cord. Although anti-hypersensitive, anti-edematous and anti-pyretic actions of MCN or PMF, in combined treatments PMF exposure decreased the anti-hyperalgesic and anti-allodynic actions of MCN treatment. These may be associated with decreases in IL-4 and IL-10 levels and an increase in CCL3 level of spinal cord tissues. CONCLUSION: Present findings support that MCN or PMF has anti-inflammatory properties duo to the down-regulating central microglial and/or peripheral inflammatory markers. Our data showed here, for the first time, PMF exposure may suppress the anti-hypersensitive actions of MCN by modulating microglia function/phenotype and microglial markers.

Prostaglandin E2 Reverses the Effects of DNA Methyltransferase Inhibitor and TGFB1 on the Conversion of Naive T Cells to iTregs.

Naturally occurring regulatory T cells (nTregs) are produced under thymic (tTregs) or peripherally induced (pTregs) conditions in vivo. On the other hand, Tregs generated from naive T cells in vitro under some circumstances, such as treatment with transforming growth factor-ß (TGFB), are called induced Tregs (iTregs). Tregs are especially characterized by FOXP3 expression, which is mainly controlled by DNA methylation. nTregs play important roles in the suppression of immune response and self-tolerance. The prostaglandin E2 (PGE2) pathway was reported to contribute to regulatory functions of tumor-infiltrating nTregs. In this study, we examined whether PGE2 contributes to the formation of iTregs treated with TGFB1 and 5-aza-2'-deoxycytidine (5-aza-dC), which is a DNA methyltransferase inhibitor. We found that the protein and gene expression levels of FOXP3 and IL-10 were increased in 5-aza-dC and TGFB1-treated T cells in vitro. However, the addition of PGE2 to these cells reversed these increments significantly. In CFSE-based cell suppression assays, we demonstrated that PGE2 decreased the suppressive functions of 5-aza-dC and TGFB1-treated T cells.

Anti-inflammatory properties of Liposome-encapsulated clodronate or Anti-Ly6G can be modulated by peripheral or central inflammatory markers in carrageenan-induced inflammation model.

Overproduction of inflammatory markers by immune cells, such as macrophages and neutrophils, is one of the main reasons for many inflammatory conditions and inhibiting or suppressing of their production by cell depletion may provide new therapeutic targets or approaches to prevent a variety of inflammatory conditions. In this study, we examined the possible effects of anti-Ly6G-mediated systemic neutrophil depletion and liposome-encapsulated clodronate (LEC)-mediated systemic macrophage depletion on the inflammatory signs (thermal hyperalgesia, mechanical allodynia, oedema and fever) and measured the levels of various inflammation markers (tumour necrosis factor-α (TNF-α), interleukins (IL)-1ß, IL-4, IL-10, macrophage inflammatory protein-1 alpha (MIP-1α/CCL3) and myeloperoxidase (MPO) in paw and spinal cord tissues in carrageenan (CG)-induced hindpaw inflammation model in rats. CG injection into the paw caused inflammation characterized by redness, swelling, heat and pain hypersensitivities. Anti-Ly6G or LEC significantly ameliorated the pain behaviours, and decreased the oedema and fever. Efficacies of anti-Ly6G or LEC on inflammatory responses changed depend on the degree of inhibition in inflammatory markers of inflamed paw or spinal cord. Anti-inflammatory properties of anti-Ly6G or LEC suggest that macrophages and/or neutrophil-mediated inflammatory cascade in inflamed site and spinal cord which can play key roles in inflammatory pain responses. These systemic or peripheral inflammatory mediators may be therapeutic targets in the treatment of many inflammatory conditions and related various diseases.

Evaluation of the Relationship Between Microalbuminuria and Urine Ischemia-Modified Albumin Levels in Patients with Diabetic Nephropathy.

INTRODUCTION: Ischemia-modified albumin (IMA) is a marker which can be associated with oxidative stress in various ischemic and non-ischemic processes. Oxidative stress plays roles in diabetes mellitus, its complications and pathogenesis. Serum IMA levels are examined in various clinical events. However, urine IMA levels have not yet been evaluated in diabetic patients. In this study, we aim to examine the relationship between metabolic features and urine microalbuminuria levels of diabetic patients and their urine IMA levels. MATERIALS AND METHODS: There were totally 50 type 2 diabetic patients in the study at the Mevlana University Hospital. Patients with cerebrovascular disease, acute myocardial infarction, hemodialysis patients with end stage chronic renal failure, pulmonary embolism, and malignant disease were excluded from the study. Metabolic features, urine IMA levels and cardiological parameters of patients were evaluated. RESULTS: Mean age of patients was 59 ± 9 years, 20 of them (40%) were male and 30 of them (60%) were female. There were six patients with albuminuria value of <0.03 mg/g (normal), there were 39 patients with microalbuminuria value of 0.03-0.3 mg/g and there were five patients with macroalbuminuria of >0.3 mg/g. According to the analysis of patients with microalbuminuria (n = 39), there was no correlation between IMA levels and numerical demographic data, albuminuria, glucose, HbA1c, lipid profile, creatinine, uric acid, hematological parameters. DISCUSSION: Conclusively, there was no relationship between urine IMA levels and microalbuminuria related to the diabetic nephropathy. These findings can be associated with urinary excretion mechanisms of IMA.

Cu (II) Chemosensor Based on a Fluorogenic Bodipy-Salophen Combination: Sensitivity and Selectivity Studies.

A new fluorescent chemosensor (Bodipy-S) derived from Bodipy and Salophen was developed. After the characterization of all compounds, the behavior of the chemosensor Bodipy-S toward p, d and f block-metal ions was investigated by UV-vis and fluorescence spectroscopy. This chemosensor can selectively detect to Cu (II) in methanol-aqueous solution based on chelation enhanced fluorescence (CHEF) and it almost exhibit to a fluorescence quenching effect with 20-fold. The binding constant of the fluorophore was interpreted by using of the Stern-Volmer method and the complex stoichiometry was defined by using Job's plot. Moreover, the effect of pH was performed by the fluorescence intensities of Bodipy-S in presence of Cu(II) ions. The chemosensor can be successfully used to the detection of Cu(II) in most areas.

Oxidative stress parameters and serum magnesium levels in patients with seasonal allergic conjunctivitis.

OBJECTIVE: To evaluate oxidative stress parameters and serum magnesium (Mg) levels in patients with seasonal allergic conjunctivitis (SAC) during the pollen season. METHODS: This observational cross-sectional study involved 35 patients with SAC without any other ocular and systemic diseases, and 38 consecutive, age- and sex-matched healthy subjects. Serum malondialdehyde (MDA), adjusted ischemia modified albumin (IMA), and Mg levels were quantified, and the results were compared between the groups. RESULTS: No significant differences were found between the groups with respect to age (p = 0.416) and sex (p = 0.362). Serum MDA and adjusted IMA levels of the subjects with SAC (69.54 ± 7.71 µM and 0.74 ± 0.39 ABSU) were significantly higher than the control group (64.61 ± 5.89 µM and 0.57 ± 0.19 ABSU) (p = 0.002 and p = 0.025, respectively). There was no significant difference for serum Mg levels between the groups (p = 0.177). CONCLUSION: We demonstrated higher levels of oxidative stress parameters in patients with SAC compared to the control group, which imply a possible role of oxidative stress in the pathogenesis of SAC.

In vitro interferon γ improves the oxidative burst activity of neutrophils in patients with chronic granulomatous disease with a subtype of gp91phox deficiency.

AIM OF THIS STUDY: Chronic granulomatous disease (CGD) is a genetically heterogeneous primary immunodeficiency caused by a defect in phagocyte production of oxygen metabolites, and resulting in infections produced by catalase-positive microorganisms and fungi. Interferon γ (IFN-γ) has a multitude of effects on the immune system. Although preliminary studies with CGD patients on treatment with IFN-γ showed that it enhanced phagocytosis and superoxide production, ongoing studies did not reveal a significant increase of this function. Here we investigated the oxidative capacity of phagocytes in different subtypes of CGD patients on treatment with IFN-γ in vitro. MATERIAL AND METHODS: Fifty-seven patients with CGD from 14 immunology centres were enrolled to our multi-centre study. Twenty-one patients were studied as controls. Oxidative burst assay with dihydrorhodamine 123 (DHR) was used and the stimulation index (SI) was calculated with respect to CGD subtypes in both neutrophils and monocytes before, and then one and 24 hours after adding IFN-γ. RESULTS: Upon comparison of the SIs of the patients' neutrophils before in vitro IFN-γ at hour 0, and after adding IFN-γ at hour 1 and 24 were compared, and the differences were determined between hours 0-24 and hours 1-24. This difference was especially apparent between hours 1-24. In CGD subtypes, particularly in gp91phox subtype, it was seen that, following in vitro IFN-γ, SIs of neutrophils began to increase after hour 1, and that increase became more apparent at hour 24. CONCLUSIONS: Our study showed that IFN-γ treatment may increase the oxidative bursting activity by increasing the superoxide production in neutrophils, particularly in gp91phox subtype.

The relationship between parental awareness of sexual abuse and children's skills to say "no".

INTRODUCTION: Child sexual abuse is a global and multidimensional social problem and causes devastating and permanent psychological, emotional, cognitive, behavioural, physical, sexual and interpersonal sequelae. This study examines the relationship between the ability to say "no" and parental awareness of sexual abuse in 4th grade primary school students. METHODS: The study was conducted between April 2022 and June 2022 in primary schools in the central district of a province in north-eastern Turkey. The sample consisted of 310 students enrolled in 4th grade and their parents. We collected the data through a personal information form, the Ability to Say "No" Scale for Children and the Sexual Abuse Awareness Scale for Parents. RESULTS: There was a weak positive correlation between the mean maternal scores of sexual abuse awareness and the mean scores of refusal and resistance in children (P < .05), as well as a weak positive correlation between the mean paternal scores of sexual abuse awareness and the mean scores of refusal and resistance in children (P < .05). CONCLUSION: As mothers' and fathers' awareness of sexual abuse myths and of teachings and actions to combat sexual abuse increased, the refusal of children also increased. Also, as fathers' awareness of the signs of sexual abuse increased, children's refusal increased.

Circulating endothelial cells in pediatric renal transplant recipients.

BACKGROUND: An increase in the number of circulating endothelial cells (CEC) indicates endothelial damage and the risk of cardiovascular disease. The aim of our study was to investigate the association of CEC with various clinical parameters in pediatric renal transplant recipients. METHODS: CEC, defined as CD45(-)CD146(+), were enumerated by flow cytometry from the peripheral blood of 50 pediatric renal transplant recipients and 20 healthy controls. Clinical parameters, including renal function tests, fasting blood glucose, serum cholesterol and triglyceride, cyclosporine A (CsA) (trough and 2nd-hour) and tacrolimus (tac) trough blood levels and their association with CEC numbers were analyzed. RESULTS: CEC numbers of patients were higher than those of controls (respectively, 128 ± 89 cells/ml (42-468 cells/ml), 82 ± 33 cells/ml (32-137 cells/ml), p = 0.024). There was a statistically significant negative correlation between CEC numbers and glomerular filtration rate (GFR) (r = -0.300, p = 0.012). There was also a statistically positive association between CEC numbers and transplant duration as well as cyclosporine trough level (respectively, r = 0.397, p = 0.004, r = 0.714, p = 0.004). CEC numbers in patients on tac and CsA were similar (p = 0.716). CONCLUSIONS: Our results demonstrate that renal transplant recipients with high CsA trough blood level, longer transplant duration, and lower GFR, are at greater risk of developing endothelial damage.

Evaluation of the Regulatory Role of Circadian Rhythm Related Long Non-Coding RNAs in ADHD Etiogenesis.

OBJECTIVE: ADHD is associated with increased sleep problems and circadian rhythm disturbances. This study aimed to examine ADHD patients and healthy controls in terms of chronotypic features and expression levels of CLOCK, PER1, lncRNA HULC, lncRNA UCA1. METHOD: Eighty-three children were included (43 ADHD). Conner's Parent Rating Scale-Revised Short Form, Childhood Chronotype Questionnaire, Children's Sleep Disorders Scale were administered. Gene expression levels were studied from peripheral blood. RESULTS: Evening chronotype, sleep initiation/maintenance disorder, sleep-wake transition disorder, excessive sleepiness disorder were higher in the ADHD group compared to the controls in the scales reported by the parents. Expression levels of all examined genes were statistically significantly higher in the ADHD group. There was no significant relationship between genes and sleep parameters in the ADHD group. CONCLUSION: Our study provides the first evidence that lncRNA HULC and lncRNA UCA1 might have a role in the etiology of ADHD.

Evaluation of real-time PCR and flow cytometry efficiency in rapid detection of carbapenemase-producing Enterobacteriales.

INTRODUCTION: Infections due to carbapenem-resistant Enterobacteriales, which have increased worldwide in recent years, cause concern. This study aimed to rapidly detect carbapenemase gene region by using flow cytometry in Enterobacteriales isolates and to evaluate its efficiency and susceptibility by comparing it with polymerase chain reaction (PCR). METHODOLOGY: In the study, 21 isolates obtained from the blood cultures of patients hospitalized in intensive care units and found to intermediate or resistant to at least one carbapenem in the automated system, and 14 isolates belonging to the carbapenem-susceptible Enterobacteriales family were included. Carbapenemase gene regions were investigated by PCR after their susceptibility was determined by disk diffusion method. Bacterial suspensions were treated with meropenem + specific carbapenemase inhibitors (EDTA or APBA) and Temocillin and stained with thiazole orange (TO) and propidium iodide (PI) to show dead/live cell differentiation. Dead/live cell percentages were calculated after reading on the flow cytometer device. RESULTS: In the ROC analysis of the flow cytometry method, the cut-off value, specificity, and susceptibility of PI staining rates for meropenem were found as 14.37%, 100%, and 65%, respectively. It was found that the flow cytometry method was well-compatible with PCR in the detection of the carbapenemase gene region. CONCLUSIONS: Flow cytometry will continue to be a promising method for the detection of antimicrobial susceptibility and resistance due to its rapid analysis of many cells and its high compatibility with PCR results.

Total antioxidant capacity, hydrogen peroxide, malondialdehyde and total nitric oxide concentrations in patients with severe persistent allergic asthma: its relation to omalizumab treatment.

BACKGROUND: Asthma is a chronic inflammatory disease and omalizumab is indicated for moderate-to-severe persistent asthma. The results of many studies have shown that oxidative stress is involved in asthma pathogenesis. However, there is no data available to evaluate the alterations in total antioxidant capacity, hydrogen peroxide, malondialdehyde, and total nitric oxide concentrations. OBJECTIVES: The objective was to determine whether treatment with omalizumab in severe allergic asthma influences systemic levels of oxidative stress markers. METHODS: The first group of 14 patients included 6 male and 8 female subjects with severe persistent asthma, having a mean age of 42.4 years. The second group included 14 newly diagnosed allergic asthma patients with a mean age of 43.8 years. All patients were followed in the Immunology and Allergy Clinic of the Antalya Education and Training Hospital and were evaluated by clinical status. A third group of 14 age-sex matched healthy controls were also included. Serum samples were collected and stored at -70 degrees C until use for the determination of total antioxidant capacity, hydrogen peroxide, malondialdehyde, and total nitric oxide concentrations. Serum IgE levels, ANA (antinuclear antibody), RF (rheumatoid factor), hepatitis markers, C3, C4, and eosinophil levels were evaluated in all patients. All assays were carried out in duplicate. RESULTS: The mean IgE levels were as follow: Group I: 459.785 IU/mL; Group II: 124.8 IU/mL, and Group III: 39.88 IU/mL. Total antioxidant capacity levels of Group IB, group II, and group III were lower than the IA group. Total antioxidant capacity levels of groups II and III were higher than in group IB. Hydrogen peroxide concentrations in group IB were lower than in group IA, while concentrations in group II were higher than in group IB. The malondialdehyde concentration of group IB was lower than in all other groups. The malondialdehyde concentration of group III was higher than all other groups. The malondialdehyde concentration of group II was lower than in group III. The total nitric oxide level of group IB was lower than all other groups. The total nitric oxide level of group III was higher than all other groups, while that of group II was higher than for both groups IA/IB. CONCLUSIONS: To monitor the omalizumab treatment efficacy in severe allergic asthma patients, total antioxidant capacity, hydrogen peroxide, malondialdehyde, and total nitric oxide concentrations might be new markers.

Pulsed magnetic field treatment ameliorates the progression of peripheral neuropathy by modulating the neuronal oxidative stress, apoptosis and angiogenesis in a rat model of experimental diabetes.

OBJECTIVE: The present study aimed to investigate the possible anti-neuropathic effects of daily pulsed magnetic field treatments (PMF) in streptozotocin (60 mg/kg) induced 4 weeks diabetic (type-1) wistar rats (6-8 months). MATERIALS AND METHODS: Body mass, blood glucose and thermal and mechanical sensations were evaluated during the PMF or sham-PMF in diabetic or non-diabetic rats (n = 7/group). After the measurements of motor nerve conduction velocities (MNCV), the levels of several biomarkers for oxidative stress, apoptosis and angiogenesis in spinal cord and sciatic nerve were measured. RESULTS: PMF for 4 weeks significantly recovered the MCNV (96.9% and 63.9%) and almost fully (100%) restored to the latency and threshold. PMF also significantly suppressed the diabetes induced enhances in biochemical markers of both neuronal tissues. CONCLUSIONS: Findings suggested that PMF might prevent the development of functional abnormalities in diabetic rats due to its anti-oxidative, anti-apoptotic and anti-angiogenic actions in neuronal tissues.

The effect of a psychoeducation program based on the rational emotional behavioral approach in individuals with multiple sclerosis diagnosis: A randomized controlled trial.

PURPOSE: This study aimed to examine the effects of a psychoeducational program on the cognitive emotion regulation levels of individuals with multiple sclerosis (MS). DESIGN AND METHOD: This study followed a randomized, controlled quasi-experimental design. A questionnaire including descriptive characteristics and disease-related characteristics and the Cognitive Emotion Regulation Questionnaire were used in the data collection. FINDINGS: The difference between the pretest-posttest and follow-up test mean scores of the patients in the experimental group was significant (p < 0.05). CONCLUSION: The psychoeducation program based on the rational emotional behavioral approach increased the use of cognitive emotion regulation strategies in individuals with MS.

DNA methylation or histone modification status in metastasis and angiogenesis-related genes: a new hypothesis on usage of DNMT inhibitors and S-adenosylmethionine for genome stability.

Metastasis is a leading cause of mortality and morbidity in cancer. This process needs angiogenesis. The biology underlying cancer, metastasis, and angiogenesis has been investigated so as to determine the therapeutic targets. Invasive and metastatic cancer cells have undergone numerous genetic and epigenetic changes, manifested by cytoskeletal changes, loss of adhesion, and expression of proteolytic enzymes that degrade the basement membrane. Additionally, in endothelial cells, some epigenetic modifications occur during the formation of angiogenesis. Researchers have used some methylation inhibitors, histone deacetylase inhibitors, or methylating agents (such as S-adenosylmethionine, SAM) against cancer and angiogenesis. Although they are effective to beat these diseases, each one results in differentiation or changes in genome structure. We review epigenetically modified genes related with angiogenesis and metastasis in cancer and endothelial cells, and suggest a new proposal. This hypothesis has discussed the importance of the usage of DNA methylation inhibitors together with SAM to prevent tumor progression and genome instability or changes resulting in additional diseases.

Inhibition of angiogenesis by S-adenosylmethionine.

Metastasis is a leading cause of mortality and morbidity in cancer. One of the steps in metastasis process is the formation of new blood vessels. Aberrant DNA methylation patterns are common in cancer cells. In recent studies, S-adenosylmethionine (SAM), which is a DNA methylating agent, has been found to have inhibitory effects on some carcinoma cells in vivo and in vitro. In the present study, we have used SAM to investigate whether it is effective against angiogenesis in vitro. Our results have shown that SAM can reduce the formation and organization of capillary-like structures of endothelial cells in tumoral environment. Besides, we have found SAM can block endothelial cell proliferation and the migration of cells towards growth factors-rich media. In conclusion, our study suggests that SAM may be used against angiogenesis as a natural bio-product.

KNK437, a benzylidene lactam compound, sensitises prostate cancer cells to the apoptotic effect of hyperthermia.

Hyperthermia is known to serve as a powerful tool in the treatment of prostate cancer which is commonly diagnosed in men. Quercetin and KNK437, Hsp70 inhibitors, play an important role in blocking thermotolerance in some cancer cells. In the present study we investigated the effects of KNK437 and quercetin on the acquisition of thermotolerance and heat-induced apoptosis. Also, it was examined whether the possible mechanism triggering apoptotic pathway included caspase-3 activation in prostate cancer cells. For this purpose, PC-3 and LNCaP cells were treated with hyperthermia following pretreatment with or without KNK437 or quercetin. Thermotolerance was investigated by colony formation assay in these cells, while Hsp70 mRNA levels were measured by real time RT-PCR. Sandwich ELISA was used for detection of Hsp70 protein levels. Apoptosis was detected by flow cytometric annexin V binding assay and by western blot analysis of procaspase-3 and cleaved poly (ADP-ribose) polymerase levels. In our study, KNK437 and quercetin inhibited thermotolerance in a dose-dependent manner in PC-3 cells. KNK437 and quercetin decreased heat-induced accumulation of Hsp70 mRNA and protein in PC-3 and LNCaP cells. KNK437 and quercetin pretreatment enhanced the apoptotic effect of hyperthermia in both cells. We found that KNK437 was more effective than quercetin in inducing apoptotic cell death, activation of caspase-3, and cleavage of PARP in prostate cancer cells. We suggest that KNK437 is a useful agent for enhancing the efficiency of hyperthermic therapy which has less toxic side-effects in prostate cancer.

Lipid peroxidation and parathyroid hormone influence the cytosolic calcium levels of erythrocytes in peritoneal dialysis patients.

BACKGROUND: The aim of this study was to examine the alterations in calcium and lipid peroxidation in red blood cells (RBCs) and serum samples of continuous ambulatory peritoneal dialysis (CAPD) patients. We also investigated the relationship between parathyroid hormone (PTH) and calcium homeostasis in this study. METHODS: For this purpose, routine blood counts and blood chemistry were analyzed by standard laboratory procedures in serum samples. The concentration of TBARS was measured in erythrocytes and serum samples. RBC calcium was measured by Fura-2AM in a spectrofluorometer. RESULTS: In CAPD patients, hemoglobin, albumin, and high density lipoprotein cholesterol levels were lower, but glucose, very low density lipoprotein cholesterol, triglyceride, magnesium, PTH, sensitive C-reactive protein, and uric acid levels were higher than the controls. Thiobarbituric acid-reactive substance (TBARS) levels in RBCs and serum samples and cytosolic calcium in RBCs were all found to be significantly increased in CAPD patients compared to control subjects. Multiple regression analysis showed that RBC TBARS and serum PTH were the independent predictors of RBC calcium in our study. CONCLUSIONS: Our results confirm that oxidative stress is an important risk factor for CAPD. The results of multiple regression analysis suggest that RBC calcium was affected by both increased levels of TBARS and PTH.

CCT3 suppression prompts apoptotic machinery through oxidative stress and energy deprivation in breast and prostate cancers.

Mediated by chaperon proteins, protein misfolding plays a crucial role in cancer pathogenesis. Chaperonin Containing TCP1 Subunit 3 (CCT3) is one of eight subunits forming eukaryotic chaperons that catalyzes correct folding of the proteins employed in cell division, proliferation, and apoptosis pathway. Moreover, CCT3 expression increases responsively with carcinogenesis. However, how CCT3 drives the cancerous process has not been documented. Here we probed the mechanistic and functional interactions between CCT3 and apoptotic pathways and cell stressors. First, we profiled CCT3 expression levels of different 16 cell lines and found that CCT3 expression levels of CRL-2329 and PC3 were significantly increased. Then, we suppressed CCT3 levels in CRL-2329 and PC3 lines by miR-24-3p, miR-128-3p, and miR-149-5p mimics, and measured apoptotic response of the cell lines to the knockdown of CCT3 by acridine orange/ethidium bromide and Annexin V/PI staining, cell-cycle and mitochondria membrane potential (MMP) analyses, intracellular reactive oxygen species (ROS) measurement and analysis of expression levels of the apoptotic genes. After having suppressed CCT3, the cell cycle was arrested in the G0/G1 phase, MMP was impaired, and the intracellular ROS level was increased. These signs of apoptotic flux were corroborated by morphological images, statistically enhanced expression levels of the apoptotic pathway modulators and intracellular free amino acids profile. The free amino acid profile, which is heavily implicated in energy metabolism and cell division, is fluctuated in the progress of canceration. Strikingly, suppressed CCT3 shifted intracellular levels of glutamine, beta-alanine, glycine, serin, asparagine and sarcosine, which are employed in energy metabolism. Consequently, miRNA-mediated CCT3 suppression spur apoptosis by unbalancing the homeostasis in intracellular ROS and the profile of free amino acids in energy metabolism. Taken together, we anticipate that miRNA-mediated CCT3 suppression might provide a "dual therapeutic strategy" through conventional cellular toxicity as well as energy withdrawal.

Neuroprotective and anti-neuropathic actions of pulsed magnetic fields with low frequencies in rats with chronic peripheral neuropathic pain.

The management of chronic peripheral neuropathic pain conditions with conventional treatments is still limited. In this present study, we aimed to determine the anti-neuropathic actions of pulsed magnetic field (PMF) treatments as a therapeutic. Effects of daily PMF treatments for 4 weeks were investigated by examining pain behaviors, hyperalgesia and allodynia, electrophysiological parameters, amplitude of compound action potential (CAP) and sciatic nerve conduction velocity (SNCV) and histopathological changes in rats with chronic constriction injury (CCI). Peripheral and central pro-inflammatory cytokines (TNF α, IL-1ß and IL-17), chemokines (CCL3 and CXCL1) and angiogenic factors (VEGF and bFGF) in sciatic nerves and spinal cord tissues were also measured for determining the possible molecular action mechanisms of PMF treatment. Hyperalgesia and allodynia were observed at the first week and lasted for 4 weeks after CCI. PMF treatments caused time-dependent anti-hyperalgesic and anti-allodynic effects. PMF treatment alleviated the histopathological consequences of CCI on sciatic nerve and significantly improved the amplitude of the CAP and SNCV. PMF treatment inhibited the pro-inflammatory molecules and promoted the anti-inflammatory cytokines in neural tissues. PMF treatment also suppressed the VEGF levels and enhanced the bFGF levels in both neural tissues. The results of the present study suggested that daily PMF treatment may have neuroprotective and anti-neuropathic pain actions in rats with CCI-induced neuropathy due to its modulating effects on neuro-inflammatory and neuro-angiogenic mediators in central and peripheral neural tissues.