Design, synthesis, molecular modeling, in vitro evaluation of novel piperidine-containing hydrazone derivatives as cholinesterase inhibitors.

In an effort to develop new and effective therapeutic agents for Alzheimer's disease, a series of hydrazone derivatives bearing piperidine rings have been designed and synthesized. The chemical structures of the compounds were characterized by various spectroscopic techniques. In vitro antioxidant and cholinesterase activities of the compounds were evaluated. Among the compounds, N12 exhibited the most antioxidant activity in all methods (CUPRAC, FRAP, DPPH, ABTS). In vitro acetylcholinesterase (AChE) activity results of the compounds showed good IC50 values between 14.124 ± 0.084 and 49.680 ± 0.110 µM were obtained (IC50 = 38.842 ± 0.053 µM for Donepezil). Among the compounds, N7 and N6 are much more effective derivatives than the standard compound donepezil with IC50 values of 14.124 ± 0.084 and 17.968 ± 0.072 µM, respectively. In vitro, butyrylcholinesterase (BChE) inhibition values of the compounds were between 13.505 ± 0.025 and 52.230 ± 0.027 µm. Among the compounds, N6 has the highest BChE inhibition with an IC50 value of 13.505 µm in the series. The cytotoxicity and AChE inhibitory activity of the compounds on SH-SY5Y cell lines were also evaluated. Kinetic studies were also performed to determine the behavior of the compounds as competitive or noncompetitive inhibitors. The binding modes of N6, which was determined to be highly effective according to in vitro analyses, with AChE and BChE were investigated using molecular docking studies, and the stability of the complexes was determined by molecular dynamics simulations. These findings indicated that AChE and BChE enzymes maintained their overall structural stability and compactness during interactions with compound N6.

Effect of platelet exosomes loaded with doxorubicin as a targeted therapy on triple-negative breast cancer cells.

Exosomes have come to the fore in drug delivery systems due to their biological-based and immune-suppressing properties. In this study, we investigated the effect of doxorubicin loading of exosomes isolated from human platelets on breast cancer.Exosomes released from ADP (1 µM)-activated platelets were isolated by the ultracentrifugation method, and their size and charge were measured with a TEM and zeta sizer. Then doxorubicin (Dox) loading into exosomes (PLT-Exo-Dox) was done by electroporation and incubated with MDA-MB-231 cells. In exosome characterization, CD62 positivity was higher in platelet pellets, while CD9 positivity was higher in released exosomes. The size of PLT-Exo and PLT-Exo-Dox was 82.02 ± 5.21 nm and 116 ± 3.73 nm, with a polydispersity index of 0.26 ± 0.04 and 0.39 ± 0.06, and the Zeta potential was - 16.45 mV and 24.07 mV, respectively. The encapsulation efficiency of the preparation was 86.02 ± 6.16%, with a drug loading capacity of 4.75 ± 0.16 µg/µg of the exosome. In MDA-MB-231 cells, PLT-Exo increased cell viability, while PLT-Exo-Dox decreased in 24 h. The Annexin-V binding level and Bax gene expression were increased in PLT-Exo-Dox and Bcl-2 gene expression was decreased. This study will shed light on the development of release systems that can be effective with chemotherapeutic agents by using exosomes released by cells in the development of personalized treatments.

Human wharton-jelly mesenchymal stromal cells reversed apoptosis and prevented multi-organ damage in a newborn model of experimental asphyxia.

In this study, the effect of applying wharton jelly mesenchymal stromal cells (WJ-MSC) isolated from the human umbilical cord tissue on the neonatal mouse model caused experimental asphyxia in mice was investigated. WJ-MSC surface markers (CD44, CD90, CD105) were characterised by immunofluorescence staining, and pluripotency genes (Nanog, Oct-4, Sox-2) were characterised by qPCR. Blood, prefrontal cortex, cerebellum, hippocampus, lung, heart, kidney, and liver tissues were analysed twenty days after subcutaneously administered WJ-MSC. WJ-MSC administration significantly decreased serum TNF-α, NSE, GFAP, and IL-6 levels in the asphyxia mice. It was determined that WJ-MSC application in tissues accelerated cell regeneration and decreased oxidative stress. In conclusion, this study showed that multiorgan damage in asphyxia could be prevented by applying WJ-MSC at an early stage. Therefore, WJ-MSC application in infants with neonatal asphyxia in the clinic may be an innovative method in the future.

Gingival crevicular fluid levels of interleukin-18 and tumor necrosis factor-alpha in type 1 diabetic children with gingivitis.

OBJECTIVE: This study aimed to evaluate the levels of interleukin-18 and tumor necrosis factor-alpha in gingival crevicular fluid of diabetic children with gingivitis. METHODOLOGY: Eighty-eight children (44 with type 1 diabetes mellitus and 44 systemically healthy) were recruited for the study. The children were divided into four subgroups based on their periodontal and systemic condition: (1) systemically and periodontally healthy children (H), (2) systemically healthy children with gingivitis (G), (3) periodontally healthy children with T1DM (T1DM + H), and (4) children with T1DM and gingivitis (T1DM + G). The plaque index, gingival index, probing pocket depth, and GCF volume were recorded. The IL-18 and TNF-α levels in GCF were determined by ELISA. RESULTS: The clinical periodontal parameters, GCF IL-18 level, and TNF-α level were similar between diabetic and systemically healthy children (p > 0.05). The gingivitis subgroups had a significantly higher GI, PI, PPD, GCF volume, and TNF-α total amounts than the H subgroups (p < 0.0001). The IL-18 concentrations in the gingivitis subgroups were significantly lower than in the periodontally healthy subgroups. CONCLUSIONS: In diabetic children with good metabolic control, T1DM did not affect the GCF levels of IL-18 and TNF-α in the presence of gingivitis. However, increased GCF TNF-α levels in children with gingivitis confirm that TNF-α is closely related to gingival inflammation. CLINICAL RELEVANCE: Type 1 diabetes mellitus is not associated with GCF interleukin-18 and tumor necrosis factor-alpha levels in children with gingivitis.

Quercetin protects mouse oocytes against chromium-induced damage in vitro and in vivo.

BACKGROUND: Chromium (Cr) is a naturally-occurring element that is used in various fields of industry. Humans may be exposed to hexavalent chromium [Cr(VI)], which is one of the stable valence states of the chromium through contaminated soil, air, and water. Exposure to Cr(VI) through contaminated drinking water, soil and air causes various cancers and also fertility problems in animals and humans. Quercetin (QCT), a common flavonoid compound, has numerous biological effects as an antioxidant and free radical scavenger, but its function and mechanisms in reproductive processes in various species remain unclear. This study aims to determine the chromium effects on mice oocyte quality and the ameliorative effect of QCT in both in vitro and in vivo experimental models. METHODS: For the in vitro experiment, oocytes were collected and divided into the control, sham, QCT-treated, Cr(VI) (potassium dichromate), and treatment [Cr(VI)+QCT] groups. Collected oocytes were cultured in maturation medium with or without 10 µM quercetin and 10 µM Cr(VI) for 14 h based on the defined experimental design. For the in vivo experiment, the mice were randomly divided into the control, sham, QCT-treated, Cr(VI), and Cr(VI) + QCT groups. Control and sham mice received regular drinking water and diet. Cr(VI) group received Cr(VI) (50 ppm in drinking water) and Cr(VI) + QCT group received 50 ppm Cr(VI) with QCT (20 mg/kg body wt, through i.p) for a period of 21 days and then oocytes were collected and cultured for 14 h for in vitro maturation. For both experiments, at the end of the culture period, we examined the ameliorative effect of QCT on oocyte maturation, spindle formation, ROS production, mitochondrial function, and apoptosis. RESULTS: Our in vitro and in vivo results showed that Cr(VI) disrupt the oocyte maturation and spindle formation (P < 0.001). Furthermore, we found that exposure to Cr(VI) significantly increased ROS levels and decreased mitochondrial membrane potential (P < 0.001). In addition, exposure to Cr(VI) induced early apoptosis and downregulated the Bcl-2 mRNA expression and upregulated the Caspase-3 and Bax mRNAs expression (P < 0.01). Finally, quercetin significantly restored the detrimental effects of Cr(VI). CONCLUSION: The results indicated that quercetin protects the oocytes against Cr(VI) toxicity through the suppression of oxidative stress and apoptosis. The conclusions drawn from our study's findings suggest that quercetin might be useful agent for oocyte maturation in case of possible exposure to toxic substances such as chromium.

Wharton jelly-derived mesenchymal stem cell exosomes induce apoptosis and suppress EMT signaling in cervical cancer cells as an effective drug carrier system of paclitaxel.

Mesenchymal stem cells can be obtained and multiplied from various sources and have a very high capacity to release exosomes. Exosomes are nano-sized extracellular vesicles containing biological signaling molecules. This study aimed to determine the effect of MSC-derived exosomes as a drug delivery system for paclitaxel in cervical cancer cells. In this study, human MSC were isolated from wharton jelly of umbilical cord tissue (WJ-MSC), and cells were characterized by CD44, CD90, CD105, and CD34 staining. Exosomes were released in WJ-MSC cells with serum-starved conditions for 48 hours, and particle sizes and structures were examined with zeta-sizer and TEM. In addition, exosomes CD9, CD63, and CD81 markers were checked by western blot. Paclitaxel was loaded into exosomes (Exo-PAC) by electroporation and then incubated with Hela cervical cancer cells for 24 hours. TGF-ß, SMAD, Snail, Slug, ß-catenin, Notch, Caspase-3, Caspase-9, Bax, Bcl-2 protein and gene expression levels were analyzed in Hela cells. As a result, low concentration Exo-PAC induced apoptosis, and suppressed epithelial-mesenchymal transition proteins in Hela cells. In this study, it has been demonstrated that WJ-MSCs can be used as drug delivery systems for cervical cancer if exosomes are produced scalably in the future.

On-Chip Organoid Formation to Study CXCR4/CXCL-12 Chemokine Microenvironment Responses for Renal Cancer Drug Testing.

Organoid models have gained importance in recent years in determining the toxic effects of drugs in cancer studies. Organoid designs with the same standardized size and cellular structures are desired for drug tests. The field of microfluidics offers numerous advantages to enable well-controlled and contamination-free biomedical research. In this study, simple and low-cost microfluidic devices were designed and fabricated to develop an organoid model for drug testing for renal cancers. Caki human renal cancer cells and mesenchymal stem cells isolated from human umbilical cord were placed into alginate hydrogels. The microfluidic system was implemented to form size-controllable organoids within alginate hydrogels. Alginate capsules of uniform sizes formed in the microfluidic system were kept in cell culture for 21 days, and their organoid development was studied with calcein staining. Cisplatin was used as a standard chemotherapeutic, and organoid sphere structures were examined as a function of time with an MTT assay. HIF-1α, CXCR4 and CXCL-12 chemokine protein, and CXCR4 and CXCL-12 gene levels were tested in organoids and cisplatin responses. In conclusion, it was found that the standard renal cancer organoids made on a lab-on-a-chip system can be used to measure drug effects and tumor microenvironment responses.

Elevated Melatonin Levels in Children With Attention Deficit Hyperactivity Disorder: Relationship to Oxidative and Nitrosative Stress.

Objective: The aim of this article is to measure serum antioxidant melatonin, the oxidants of nitric oxide, and malondialdehyde levels to calculate the serum oxidant-antioxidant balance based on the nitric oxide/melatonin and malondialdehyde/melatonin ratios in children with ADHD. Method: The serum melatonin, nitric oxide, malondialdehyde, and the nitric oxide/melatonin and malondialdehyde/melatonin ratios were calculated and compared between the children with ADHD (n = 103) and healthy control participants (n = 73). Results: Serum melatonin and nitric oxide levels were higher, and the nitric oxide/melatonin and malondialdehyde/melatonin ratios were lower in ADHD children than the control group. Melatonin was found to be significantly high, and the malondialdehyde/melatonin ratio was found to be significantly low in children with a positive ADHD family history. Conclusion: The serum oxidant-antioxidant balance was impaired in children with ADHD. Within the ADHD group, higher melatonin levels were determined in the children with a positive family history.

The effect of hypothermia in a rat testicular torsion/detorsion model.

BACKGROUND: Testicular torsion is an emergent condition. The protective effect of medical hypothermia in ischemia/reperfusion injury is well defined. OBJECTIVES: To evaluate the late results of hypothermia through a rat testicular torsion/detorsion model compatible with human testicular torsion. STUDY DESIGN: Rats were divided into 5 groups (n = 7): (1)Sham (S) group, (2)T/D group: right testis was torted for 1-h, (3)T/D + H30 group: hypothermia at 4 °C was applied for 30 min before detorsion, (4)T/D + H90 group: hypothermia at 4 °C was applied for a total of 90 min (30 min before and 1-h after detorsion), (5)H group: hypothermia at 4 °C was applied to right testis for 90 min. Testicular diameters at preoperative period and 8th postoperative week were measured. Biochemically, MPO, NO, 3-NT and 4-HNE in testicular tissue and serum levels of NO, PGF 2α, 3-NT, 8-OHdG and 4-HNE were studied. Histopathologic examination and TUNEL assay were also performed. RESULTS: Biochemical and macroscopical parameters of both T/D + H30 and T/D + H90 groups were statistically different from group T/D with respect to protective effects of hypothermia. Johnsen's score was also statistically different in group T/D + H90. DISCUSSION: Hypothermia can easily be applied with ice bags both in perioperative period. This is the first study evaluating the effect of hypothermia applied postoperatively. Tissue level of protein oxidation marker (3-NT) and serum levels of DNA damage (8-OHdG), lipid peroxidation (4-HNE), protein oxidation (3-NT) and oxidative stress (PGF-2α) markers were measured for the first time. CONCLUSIONS: Hypothermia has been shown to be macroscopically, biochemically and histopathologically beneficial in the long-term experimental testicular torsion model.

Serum Copper and Zinc Levels in Patients with Endometrial Cancer.

The aim of the present study was to evaluate serum concentrations of copper (Cu) and zinc (Zn), in relation with metabolic profile and clinicopathologic features of patients with endometrial cancer. A total of 47 women with endometrial cancer and 45 controls were eligible for the study. Clinicopathologic features and metabolic profile as well as serum copper and zinc levels were evaluated in each subject. Patients with endometrial cancer (Cu mean 3.72 ± 2.15 mg/L, median 3.54 [0.41-9.16] mg/L and Zn mean 1.83 ± 0.71 mg/L, median 1.77 [0.71-4.02] mg/L) exhibited lower Cu and Zn levels than those of controls (Cu mean 6.06 ± 1.79 mg/L, median 6.32 [2.95-9.05] mg/L and Zn mean 2.48 ± 0.89 mg/L, median 2.23 [1.23-4.54] mg/L) (p < 0.001). Cu/Zn ratio was also higher (0.85 ± 1.96 vs. 2.57 ± 0.73) in controls as compared with patients with endometrial cancer. While Cu levels showed no significant correlation with age, body mass index, gravidity, and parity, a positive correlation was found between Zn levels and parity. When cancer patients were evaluated on their own, both Cu and Zn levels showed positive correlation with age. Additionally, the cancer patients with myometrial invasion > 1/2 exhibited lower Cu levels compared with the cancer patients with myometrial invasion < 1/2. The data of the present study suggested that women with endometrial cancer are characterized by altered serum Cu and Zn levels as compared with controls. Imbalance of these trace element levels might be associated with endometrial cancer among Turkish patients.

Ozone Partially Decreases Axonal and Myelin Damage in an Experimental Sciatic Nerve Injury Model.

AIM: To investigate the effects of ozone in experimental acute sciatic nerve injury. MATERIAL AND METHODS: Twenty-eight male rats were divided into four groups (n = 7): control (C), ozone (O), injury (SNI), and treatment with ozone after injury (SNI + Ozone). Sciatic nerve injury was generated by compressing the right sciatic nerve for 90 s using a Yasargil aneurysm clip in groups SNI and SNI + Ozone. A 70 µg/ml concentration of ozone was given four times (once a day at 1st, 24th, 48th, and 72th h) at a dose of 0.5 mg/kg to groups O and SNI + Ozone after injury by an intraperitoneal injection. Nerve conduction velocities of all rats were measured by in vivo electrophysiological tests at the end of the day 4. Then, plasma malondialdehyde, total oxidant and antioxidant status were measured and also axonal and myelin changes in sciatic nerves of histopathological examination were performed. The data were analyzed by Kolmogorov Smirnov test, Mann-Whitney U-test, and Chi square test. p <.05 was considered statistically significant. RESULTS: The proximal and distal latency difference were higher and nerve conduction velocity were lower in SNI group than C and O groups, and the myelin structure was found to be broken in group SNI compared to groups C and O. However, the amplitude of the compound action potential, the nerve conduction velocity were significantly higher in group SNI + Ozone than in group SNI. Moreover, myelin injury was significantly lower in group SNI + Ozone compared to group SNI. Total oxidant status in group SNI was significantly higher than in groups C, O, and SNI + Ozone. But, total antioxidant status in group SNI was significantly lower than in groups C, O, and SNI + Ozone. CONCLUSION: This study showed that the administration of ozone at a dose of 0.5 mg/kg after peripheral nerve injury in rats reduces myelin and axonal injury.

The association between clinically suspect arthralgia and adipokines in obese patients.

OBJECTIVE: Obesity is a moderate low-grade chronic inflammatory condition. The cause of low-grade inflammation in obese patients who have clinically suspect arthralgia (CSA) may be the subject of debate in clinical practice. Our aim is to determine whether inflammation is associated with obesity or rheumatic disease, and the association between leptin, chemerin, visfatin and inflammatory markers in obese patients with/without musculoskeletal symptoms. METHODS: Seventy-four obese patients who admitted to our rheumatology clinic with CSA were enrolled. The control group consisted of 40 obese patients who have no rheumatic symptoms. Body mass index (BMI) was calculated in kg/m2 with body weight ratio to height squared, and obesity was defined as BMI 30 or above. Age, gender, BMI, waist and hip circumferences, waist-to-hip ratio (WHR), erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6), interleukin-1 beta (IL-1ß), leptin, chemerin, and visfatin were evaluated. The relationship between all parameters was assessed by Spearman correlation, Wilcoxon Signed-rank, and paired t-tests. RESULTS: There were no significant differences for age, gender, ESR and CRP between obese patients with CSA and control group. The mean TNF-α, IL-1ß, IL-6 concentrations were 60.8 pg/mL, 39.9 pg/ml, and 26.2% in obese patients with CSA, respectively. ESR, CRP, TNF-α, IL-6, and IL-1ß concentrations were higher in these patients compared to obese patients without any rheumatic symptoms. The mean WHR and waist circumference were 0.8±0.1 and 107.1±13.4 cm, respectively in patients with CSA. IL-6 correlated with WHR and waist circumference, positively. There were significant differences for adipokines such as chemerin, visfatin, but not for leptin between both group. Moreover, a significant correlation was found between pro-inflammatory cytokines and visfatin, chemerin. CONCLUSION: Visfatin and chemerin correlated with inflammation and may be useful indicators of undifferentiated inflammatory arthritis in obese patients with CSA.

Evaluation of Oxidant-Antioxidant Balance in Children with Atopic Dermatitis: A Case-Control Study.

BACKGROUND: Increased reactive oxygen species (ROS) and oxidative stress (OS) has been reported in many allergic and inflammatory skin diseases, including urticaria, psoriasis, and atopic dermatitis (AD). Melatonin is a hormone secreted from the pineal gland and is a potent antioxidant. OBJECTIVE: The aim of the study was to measure serum antioxidant melatonin, oxidants of nitric oxide (NO), and malondialdehyde levels to calculate the serum oxidant-antioxidant balance based on the NO/melatonin and malondialdehyde/melatonin ratios and to determine the correlation with the disease severity in children with AD. METHODS: Seventy-three children with AD and 67 healthy controls were included in the study. The clinical diagnosis of AD was based on the diagnostic criteria of Hanifin-Rajka. The severity of AD was evaluated by the scoring AD (SCORAD) index, and atopy was determined by skin prick tests (SPTs) with commercial extracts. The OS-related parameters of serum melatonin, NO, malondialdehyde, and the NO/melatonin and malondialdehyde/melatonin ratios were calculated and compared with the results of healthy controls. RESULTS: Serum melatonin levels were higher (p < 0.0001) and serum NO levels and the NO/melatonin and malondialdehyde/melatonin ratios were lower in children with AD than in healthy controls (p = 0.045, p < 0.0001, p < 0.0001, respectively). There was no difference between children with AD and healthy controls in terms of serum malondialdehyde levels (p = 0.119). Serum melatonin levels were significantly lower in severe AD than in mild AD (p = 0.012). However, in terms of serum melatonin levels, there was no difference between mild and moderate AD (p = 0.742) and moderate to severe AD (p = 0.301). There was no significant difference in serum NO and malondialdehyde levels and NO/melatonin and malondialdehyde/melatonin ratios among children with mild, moderate, and severe AD (p > 0.05). A negative correlation was found between serum melatonin levels and the SCORAD index (r = -0.252, p = 0.031), and a positive correlation was found between NO/melatonin and malondialdehyde/melatonin ratios (r = 0.511, p < 0.0001). There was no statistically significant relationship between age (≤24 or >24 months), disease duration (≤6 or >6 months), and sex for the OS-related parameters (p > 0.05). CONCLUSION: The serum oxidant-antioxidant balance was impaired in children with AD. Serum melatonin levels were higher in children with AD; however, this was negatively correlated with disease severity. Serum NO levels and NO/melatonin and malondialdehyde/melatonin ratios were lower in children with AD than in healthy controls. Melatonin might be used as a promising antioxidant to evaluate disease severity in children with AD. Thus, further studies are needed to clarify the role of melatonin in AD pathogenesis.

Boric acid reduces axonal and myelin damage in experimental sciatic nerve injury.

The aim of this study was to investigate the effects of boric acid in experimental acute sciatic nerve injury. Twenty-eight adult male rats were randomly divided into four equal groups (n = 7): control (C), boric acid (BA), sciatic nerve injury (I), and sciatic nerve injury + boric acid treatment (BAI). Sciatic nerve injury was generated using a Yasargil aneurysm clip in the groups I and BAI. Boric acid was given four times at 100 mg/kg to rats in the groups BA and BAI after injury (by gavage at 0, 24, 48 and 72 hours) but no injury was made in the group BA. In vivo electrophysiological tests were performed at the end of the day 4 and sciatic nerve tissue samples were taken for histopathological examination. The amplitude of compound action potential, the nerve conduction velocity and the number of axons were significantly lower and the myelin structure was found to be broken in group I compared with those in groups C and BA. However, the amplitude of the compound action potential, the nerve conduction velocity and the number of axons were significantly greater in group BAI than in group I. Moreover, myelin injury was significantly milder and the intensity of nuclear factor kappa B immunostaining was significantly weaker in group BAI than in group I. The results of this study show that administration of boric acid at 100 mg/kg after sciatic nerve injury in rats markedly reduces myelin and axonal injury and improves the electrophysiological function of injured sciatic nerve possibly through alleviating oxidative stress reactions.