Biologically active sodium pentaborate pentahydrate and Hypericum perforatum oil loaded poly vinyl alcohol: chitosan membranes.

In this study, sodium pentaborate pentahydrate (NaB) and Hypericum perforatum (HP) oil were incorporated into polyvinyl alcohol (PVA) and chitosan (CH) polymer blend to obtain membranes by solution casting method. In order to see the synergistic effects of NaB and HP oil on the biological and physical properties of the membranes NaB and HP oil were incorporated into membrane matrix in different ratios. Fourier-transform infrared spectroscopy (FTIR) results showed that no significant bond formation between the bioactive components and the PVA:CH matrix. According to mechanical test results, Young's Modulus and elongation at break decreased from 426 MPa to 346 MPa and 52.23 % to 15.11 % for neat PVA:CH membranes and NaB and HP oil incorporated PVA:CH (PVA:CH@35NaB:HP) membranes, respectively. Antimicrobial activity tests have shown the membranes were over 99 % effective against Escherichia coli, Staphylococcus aureus, and Candida albicans, underlining their potential for infection control. Cytocompatibility assay performed with Human Dermal Fibroblast (HDFa) cells highlight the biocompatibility of the membranes, revealing 74.84 % cell viability after 72 h. The properties of NaB and HP oil doped PVA:CH based membranes obtained from these experiments reveal the promise of a versatile membrane for applications in wound healing, tissue engineering and other biomedical fields.

Paroxetine's effect on the proinflammatory cytokine stimulation and intracellular signaling pathways in J774.2 cells.

Paroxetine is extensively utilized in the management of depressive and anxious conditions. Paroxetine works by increasing serotonin levels in nerve cells in the brain. However, limited information is available regarding the direct effects of paroxetine on macrophage cells. Macrophages are a type of leukocytes involved in the body's immune response, playing a crucial role in combating infections. The impact of paroxetine on macrophages has been explored in research, although a comprehensive understanding is still pending. This study aimed to research the potential of administering paroxetine to J774.2 macrophage cells to stimulate the release of GM-CSF, TNF-α, IL-12p40, and IL-6 cytokines. Additionally, we examined the mechanisms of action of paroxetine on the p38 signaling pathway, which is involved in cytokine production, and the PI3K pathway, which is an important mechanism in intracellular signaling. Our findings revealed that paroxetine induced an inflammatory response in macrophages by promoting cytokine synthesis in a non-lipopolysaccharide (LPS) environment. We observed that paroxetine triggered the inflammatory response through the PI3K signaling pathway while suppressing the p38 signaling pathway.

Escitalopram's inflammatory effect on the mammalian macrophages and its intracellular mechanism of action.

The majority of patients with depression are treated with antidepressant drugs that are in the serotonin reuptake inhibitor (SSRI) group. Different studies have been conducted on the effect of treatment with antidepressants on the level of pro-inflammatory cytokines. There have been studies on the effects of escitalopram, an SSRI group antidepressant, on the pro-inflammatory cytokine levels both in vivo and in vitro. The results of these studies do not overlap and therefore the escitalopram's effect on the immune system should be studied in more depth. In this study, we aimed to examine, in detail, the cytokine production amount by escitalopram treatment of the J774.2 macrophage cells and its intracellular mechanism of action by examining the PI3K and p38 pathways. As a result of our study, we observed that Escitalopram caused a significant increase in TNF-α, IL-6, and GM-CSF levels in mammalian macrophage cells, but did not induce IL-12p40 production. We observed that the p38 and PI3K pathways play a role in inflammation in the presence of Escitalopram.

Anti-inflammatory activity of bupropion through immunomodulation of the macrophages.

Depression might manifest itself with a chronic inflammation in different tissues and organs independent of the central nervous system. Psoriasis, Crohn's disease, and fibromyalgia are among these disorders accompanying the depression. The treatment options for these conditions are a combination of the anti-depressants and anti-inflammatory agents. Bupropion has been widely utilized as an anti-depressant. It has been preferred among the patients with Crohn's disease and psoriasis due to its anti-inflammatory role, as well. In this study, we aimed to decipher its target in the immune system. Macrophages were activated in the presence of LPS and increasing concentrations of the bupropion. TNF-α, IL-6, GM-CSF, and IL-12p40 cytokines' production levels were measured by ELISA to compare it to the control groups. These cytokines have been associated with the aggressive inflammation in different tissues. Moreover, p38 and PI3K proteins' phosphorylated levels were measured to examine whether bupropion acts through these pathways or not. Our results suggest that bupropion had anti-inflammatory action on the activated macrophages and its mechanism of action was partially dependent on p38 but independent of PI3K pathways.

Immunostimulatory activity of fluoxetine in macrophages via regulation of the PI3K and P38 signaling pathways.

Fluoxetine is an antidepressant drug that is heavily preferred in the cure of depression, which is from the selective serotonin reuptake inhibitor (SSRI) group. There are many reports on the effect of fluoxetine on the immune system, and its effect on the macrophage cells has never been looked at before. We aimed to demonstrate the cytokine production potential of fluoxetine antidepressant, which is widely used in the clinic, in the J774.2 cell line and its effect on PI3K and P38 pathways. The use of fluoxetine alone in J774.2 macrophage cells showed immunostimulatory properties by inducing the production of tumor necrosis factor-α (TNF-α), interleukin (IL) IL-6, IL-12p40, and granulocyte-macrophage colony-stimulating factor (GM-CSF) cytokines. It showed anti-inflammatory properties by completely stopping the production of cytokines (IL-6, IL12p40, TNF-α, and GM-CSF) at all concentrations where LPS and fluoxetine were used together. While PI3K and P38 pathways were not effective in the immunostimulatory effect in the presence of the drug agent, we found that the PI3K and P38 pathways were influenced during their anti-inflammatory activity.

Antimicrobial, antibiofilm, and anticancer potential of silver nanoparticles synthesized using pigment-producing Micromonospora sp. SH121.

Silver nanoparticles (AgNPs) have gained interest as an alternative pharmaceutical agent because of antimicrobial resistance and drug toxicity. Considering the increasing request, eco-friendly, sustainable, and cost-effective synthesis of versatile AgNPs has become necessary. In this study, green-made AgNPs were successfully synthesized using Micromonospora sp. SH121 (Mm-AgNPs). Synthesis was verified by surface plasmon resonance (SPR) peak at 402 nm wavelength in the UV-Visible (UV-Vis) absorption spectrum. Scanning electron microscopy (SEM) analysis depicted that Mm-AgNPs were in the size range of 10-30 nm and spherical. Fourier transform infrared spectroscopy (FTIR) confirmed the existence of bioactive molecules on the surface of nanoparticles. The X-ray diffraction (XRD) analysis revealed the face-centered cubic (fcc) structure of the Mm-AgNPs. Their polydispersity index (PDI) and zeta potential were 0. 284 and -35.3 mV, respectively. Mm-AgNPs (4-32 µg/mL) exhibited strong antimicrobial activity against Bacillus cereus, Enterococcus faecalis, Enterococcus hirae, Escherichia coli, Klebsiella pneumoniae, Proteus vulgaris, Pseudomonas putida, Staphylococcus epidermidis, Streptococcus pneumoniae, and Aspergillus flavus. Mm-AgNPs partially inhibited the biofilm formation in Acinetobacter baumannii, E. coli, K. pneumoniae, and Pseudomonas aeruginosa. Furthermore, results showed that low concentrations of Mm-AgNPs (1 and 10 µg/mL) caused higher cytotoxicity and apoptosis in DU 145 cells than human fibroblast cells. Based on the results, Mm-AgNPs have an excellent potential for treating infectious diseases and prostate cancer.

Anticancer potential of Origanum munzurense extract against MCF-7 breast cancer cell.

Origanum munzurense (O. munzurense) is an endemic species of Tunceli region of Turkey. In this study, we investigated in vitro anticancer effect of aqueous extract of O. munzurense (OME) on breast cancer cells (MCF-7). In vitro cytotoxic effect of OME was evaluated in MCF-7 cells by MTT assay. The wound-healing assay was used to examine the inhibition of migration. Annexin V/propidium iodide staining, cell-cycle distribution was assessed by flow cytometry for MCF-7 cells treated with OME. MTT results show that OME demonstrated in vitro cytotoxicity with 600 mg at 48 h on MCF-7. Doses of 400 µg/mL and 600 µg/mL OME significantly suppressed the migration rate of MCF-7 cells. OME significantly decreased the percentage of live cancer cells and showed an apoptotic effect after 48 h of incubation. These results show that OME is effective against breast cancer when administered at high doses and for a long time.

Sulfoxaflor insecticide exhibits cytotoxic or genotoxic and apoptotic potential via oxidative stress-associated DNA damage in human blood lymphocytes cell cultures.

The need for foodstuff that emerged with the rapidly increasing world population made fertilizers and pesticides inevitable to obtain maximum efficiency from existing agricultural areas. Sulfoxaflor is currently the only member of the new sulfoximine insecticide subclass of nicotinic acetylcholine receptor agonists. In the study, it was aimed to determine the in vitro genetic, oxidative damage potential, genotoxic and apoptotic effects of three different concentrations (10 µg/mL, 20 µg/mL and 40 µg/mL) of sulfoxaflor insecticide in the cultures of blood lymphocytes. In this study, the single-cell gel electrophoresis (comet), Cytokinesis Block Micronuclues Test (MN test), flow cytometry and measurement of Catalase (CAT) enzyme activity were used to determine genotoxic, apoptotic effects and oxidative damage potential, respectively. It found that there is a decrease in CPBI values and Live cell numbers. It was observed an increase in late apoptotic and necrotic cell numbers, Micronucleus frequency, and Comet analysis parameters (GDI and DCP). There is a significant difference between negative control and all concentration of insecticide for Cytokinesis Block Proliferation Index (CBPI) values and late apoptotic, necrotic and viable cell counts. An increase in CAT enzyme levels was observed at 10 and 20 µg/mL concentrations compared to control., It is found that CAT enzyme activity was inhibited at concentrations of 40 µg/mL. This study is crucial as it is the first study to investigate the impact of Sulfoxaflor insecticide on peripheral blood lymphocyte cells. The genotoxic, oxidative damage, and apoptotic effects of Sulfoxafluor insecticide on the results obtained and its adverse effects on other organisms raise concerns about health and safety.

Non-canonical anti-cancer, anti-metastatic, anti-angiogenic and immunomodulatory PDT potentials of water soluble phthalocyanine derivatives with imidazole groups and their intracellular mechanism of action.

Cancer is currently a leading health issue globally. Chemotherapy is a prominent treatment method but due to undesired side effects t, there has been a need for novel less toxic approaches. Photodynamic therapy may be listed among the alternatives for efficient and potentially less detrimental applications of cancer therapy. Canonical photodynamic therapy (PDT) approach requires a light source with a specific wavelength of light, a non-toxic photosensitizer and molecular oxygen. PDT creates the desired effect by the photochemical reaction created through interaction of these components to create reactive oxygen species that will act on the cancer cells to enable anti-cancer activities. In our study we focus on non-canonical PDT application. In this approach we are not only aiming to eliminate cancer cells in the environment but also test the anti-metastatic, anti-angiogenic and possible immunomodulatory activities of the novel photosensitizers. Moreover, in our approach, we studied the intracellular pathways that are crucial for carcinogenesis, cell cycle, apoptosis, angiogenesis, metastasis and immune function to decipher the mechanism of the action for each compound. Reactive oxygen species based explanation was not valid in our study, hence it brings out a non canonical approach to PDT applications. Our results suggests that Phthalocyanine derivatives with imidazole groups can be effectively used against lung, colon, breast and prostate cancer while differentially effecting metastasis, angiogenesis, cell cycle, apoptosis and immune system cells' activities. Based on the results, PDT application of these phthalocyanine derivatives can be an effective treatment option to replace chemotherapy to minimize the potential side effects.

Photodynamic anti-inflammatory activity of azulene derivatives on mammalian macrophages and their intracellular mechanism of action.

Azulene derivatives have been studied previously as photodynamic therapy agents. They have anti-cancer, anti-microbial and anti-inflammatory activities. Together with their photodynamic activity they enable more control on their activation which aims to decrease possible side effects that have been encountered with their constitutively active drug counterparts. In our current study we focused on photodynamic anti-inflammatory activities of two azulene derivatives whose synthesis methods were described before. We found that when mammalian macrophages J774.2 cells were incubated with these two derivatives in the presence of LPS in dark conditions, these molecules had anti-inflammatory activity at their highest concentrations based on ELISA results on the pro-inflammatory cytokine levels. After light application, both derivatives exerted strong anti-inflammatory activities by substantially decreasing the TNF, IL6, GMCSF and IL12p40 cytokine production levels. When the intracellular mechanism of action for both derivatives was tested, only one of them acted through p38 and PI3K pathways whereas the other derivative did not affect either of these pathways. Our results suggest that these two azulene derivatives can be utilized as photodynamic anti-inflammatory drug candidates.

Synthesis of novel immunomodulatory 1,4-disubstituted bis-1,2,3-triazoles by using click chemistry and their intracellular mechanism of action.

In this study, six new 1,4-disubstituted bis-1,2,3-triazole compounds, N,N'-(1,2-phenylene)bis(2-(4-R-1H-1,2,3-triazol-1-yl)acetamide), were synthesized with high yield (88-96 %) by using click chemistry and their molecular structures were characterized by using NMR, FT-IR, HRMS and elemental analysis techniques. Previous studies suggest anti-inflammatory and analgesic activities for different 1,2,3-triazole derivatives and in the light of those studies we aimed to examine these novel derivatives immunomodulatory activities on the mammalian macrophages. Pro-inflammatory cytokines (TNF, IL6, GMCSF and IL12p40) secretion levels were tested in the presence of bis-1,2,3-triazole compounds when the macrophages were activated with LPS. These new derivatives were able to suppress the production of these cytokines at different levels. Intracellular phophorylated PI3K protein levels were measured due to its prominent role in inflammatory reactions. Our flow cytometry analysis results suggested that some of these compounds were partially effective through PI3K pathway. In different inflammatory and autoimmune disease settings these novel 1,2,3-triazole derivatives can be utilized as non-steroid based anti-inflammatory drug candidates.

Design, synthesis and antiproliferative activity evaluation of fluorine-containing chalcone derivatives.

A series of new chalcones containing fluoro atom at B ring have been designed, synthesized, and evaluated to be antiproliferative activity against a panel of human tumor cell lines. Some of the analogs (8, 9, 12, 45, 46 and 48) displayed powerful antiproliferative effects to certain human tumor cells, but all of them were devoid of any cytotoxicity towards the normal HEK 293. Acridine orange staining data supported that the cytotoxic and antiproliferative effects of the synthesized analogs on tumor cells are mediated through apoptosis. The compounds 12 and 46 manifested concentration-dependent antiproliferative activity in human hepatocellular carcinoma cell lines using an xCELLigence assay. The structures and antiproliferative activity relationship were further supported by in silico molecular docking study of the compounds against tubulin protein which suggests our compounds interference to cell division. Communicated by Ramaswamy H. Sarma.

Antiproliferative activity of Tamoxifen, Vitamin D3 and their concomitant treatment.

Breast cancer stands out as the most common cancer type among women throughout the world. Especially for the estrogen receptor alpha (ER α +) positive breast cancer cells Tamoxifen has been widely used as an anti-cancer agent. Tamoxifen's mechanism of action is through ER. It binds to the receptor and leads to a conformational change which eventually prevents cancer cells proliferation and survival. In our current study, we aimed to investigate the combination of Tamoxifen with Vitamin D3 to test whether this combination will enhance the anti-cancer effect of Tamoxifen on breast cancer cells in vitro. Vitamin D3 has sterol structure and this property enables it to act similar to hormones. Vitamin D Receptor (VDR) has been commonly found in different types of cancer cells including but not limited to breast and prostate cancer cells. Through this receptor Vitamin D3 acts as an anti-proliferative agent. We examined the proliferation rate, apoptosis and necrosis levels as well as cell cycle progression in MCF-7 breast cancer cell line in the presence of Vitamin D3 and Tamoxifen to compare the changes with the Tamoxifen treated group. Our results suggest that Tamoxifen was a more potent anti-cancer agent than Vitamin D3 or its combination with Vitamin D3 based on cell cycle arrest, apoptosis and cell proliferation levels. This effect in the apoptosis rate and cell cycle stage of the MCF-7 cells were in line with the changes in gene expression profile of P53, BAX and BCL-2. Our results suggest that Tamoxifen by itself is adequate enough and more potent than Vitamin D3 or its combination with Vitamin D3 as anti-cancer agent for the breast cancer cells in vitro.

Comparison of the Effects of Pulsed Electromagnetic Field and Extracorporeal Shockwave Therapy in a Rabbit Model of Experimentally Induced Achilles Tendon Injury.

Achilles tendon injuries are a common cause of complications including adhesions and tendon degeneration. As a result of these complications, the biomechanical properties are lost. Extracorporeal shockwave therapy (ESWT) and pulsed electromagnetic field (PEMF) recover the injured tendon structure; however, detailed studies of changes in tendon biomechanical properties are limited. We hypothesized that PEMF application would improve Achilles tendon biomechanical properties similar to ESWT. The curative effects of a PEMF 4-week application (15 Hz, 1 mT, 260 µs, 1 h/day) and ESWT (3 doses/28 days, 1st dose: 0.12 mJ/mm2 , 15 Hz, 300 impulses; 2nd dose: 0.14 mJ/mm2 , 15 Hz, 500 impulses; 3rd dose: 0.14 mJ/mm2 , 15 Hz, 500 impulses) on rabbits with Achilles tendon injury were investigated in terms of histopathological and biomechanical properties. The clinical feasibility of PEMF application was evaluated by comparing the results of both methods. Fifty New Zealand female rabbits were divided into two groups to be used in either biomechanical or immunohistochemical studies. Each of the two groups was further divided into five groups: C (Control), SH (Sham), TI (tendon injury), TI + ESWT, and TI + PEMF. Biomechanical evaluations revealed that maximum load, toughness, and maximum stress averages of the TI + PEMF group significantly increased (P < 0.05). When immunohistochemical images of the TI + PEMF group were compared with those of the TI group, the amount of fibrous tissue was less, the homogeneity of collagen fibers recovered, and collagen organization was more uniform. We conclude that both ESWT and PEMF are equally efficient for Achilles tendon recovery. PEMF application is effective and can be used in the clinic as a painless alternative treatment method. © 2020 Bioelectromagnetics Society.

DNA-damage and cell cycle arrest initiated anti-cancer potency of super tiny carbon dots on MCF7 cell line.

While carbon-based materials have spearheaded numerous breakthroughs in biomedicine, they also have procreated many logical concerns on their overall toxicity. Carbon dots (CDs) as a respectively new member have been extensively explored in nucleus directed delivery and bioimaging due to their intrinsic fluorescence properties coupled with their small size and surface properties. Although various in vitro/in vivo studies have shown that CDs are mostly biocompatible, sufficient information is lacking regarding genotoxicity of them and underlying mechanisms. This study aims to analyze the real-time cytotoxicity of super tiny CDs (2.05 ± 0.22 nm) on human breast cancer cells (MCF7) and human primary dermal fibroblast cell cultures (HDFa) by xCELLigence analysis system for further evaluating their genotoxicity and clastogenicity to evaluate the anti-tumor potential of CDs on breast adenocarcinoma. As combined with flow cytometry studies, comet assay and cytokinesis-block micronucleus assay suggest that the CDs can penetrate to the cell nuclei, interact with the genetic material, and explode DNA damage and G0/G1 phase arrest in cancer cells even at very low concentrations (0.025 ppm) which provide a strong foundation for the design of potentially promising CD-based functional nanomaterials for DNA-damage induced treatment in cancer therapy.

Perifosine and vitamin D combination induces apoptotic and non-apoptotic cell death in endometrial cancer cells.

Endometrial cancer is the most common cancer of the female reproductive system. Combination treatment with specific agents has been widely used as a targeted therapy for cancer. In this study, we aimed to investigate the anti-proliferative and apoptotic effects of varying concentrations of perifosine and vitamin D on the human endometrial cancer cell line (HEC-1A). HEC-1A cells were exposed to perifosine (10 µM, 30 µM), vitamin D (50 nM, 200 nM) and combinations of both for 48 h and 72 h. Monitoring of cell proliferation in a time-dependent manner was performed with the xCELLigence RTCA DP system. The levels of BCL2, BAX and P53 mRNA expression were examined using RT-qPCR. Apoptosis was determined using Annexin V, which were followed by flow cytometry analysis. Ultra-structural morphology of cells was analyzed by transmission electron microscopy (TEM) for 72 h. The anti-proliferative and apoptotic effects of the perifosine+vitamin D combination (30 µM + 200 nM at 48 h and 10 µM + 200 nM at 72 h) on HEC-1A cells were higher than in perifosine and vitamin D alone. It was observed that perifosine has increased the expression of BAX mRNA in HEC-1A cells in a dose-dependent manner. While perifosine+vitamin D combinations increased P53 mRNA expression in HEC-1A cells we did not find any significant change in BCL2, BAX mRNA expression levels. In TEM examinations of HEC-1A cells, perifosine appeared to lead autophagic cell death, whereas vitamin D caused paraptosis-like cell death and combination of perifosine+vitamin D caused apoptotic and non-apoptotic (paraptotic, autophagic and necrotic) cell death. Therefore, it is considered that the combination of both drugs in the treatment of endometrial cancer might be an alternative and effective treatment option through activating the apoptotic and non-apoptotic cell death mechanisms in cancer cells.

Effect of Imipramine on radiosensitivity of Prostate Cancer: An In Vitro Study.

Prostate cancer is the most common cancer and leading cause of cancer death for males. Imipramine (IMI), which is a tricyclic antidepressant, has also been shown to has antineoplastic effect. This study was performed to investigate the radiosensitizing effect of IMI on DU145 prostate cancer cell. Cells were divided into 4 groups. Cell index, apoptotic activity, cell cycle arrest, oxidative stress and EAG1 channel currents were determined in all groups. Our findings showed that combined treatment with IMI and radiotherapy (RAD) did not enhance radiosensitivity of DU145 cells but as unexpected finding, treatment of IMI alone was more effective in DU145 cells.

Effects of Huperzin-A on the Beta-amyloid accumulation in the brain and skeletal muscle cells of a rat model for Alzheimer's disease.

AIMS: Alzheimer's Disease (AD) is characterized by a loss of cognitive function and also the accumulation of ß-amyloid peptide (ßAP) in the brain parenchyma, which plays an important role in this disease. However, it is often also associated with the non-cognitive symptoms such as loss of muscle function (Inclusion-Body Myositis-IBM). MAIN METHODS: Sprague-Dawley rats (13 weeks-n=68) were randomly assigned into five groups: Group C: Control; Group D: d-galactose; Group O+D: Bilateral oophorectomy+d-galactose; Group O: Bilateral oophorectomy; Group O+D+H: Bilateral oophorectomy+d-galactose+Hup-A. Tissue fixation was performed with the perfusion method. The Compound Muscle Action Potential (CMAP) and mechanical muscle activity were recorded using the standard electro-biophysical techniques. Immune staining was performed with specific antibodies, and the pathological changes were examined. RNA was obtained from brain tissue samples with the Trizol Method. Then, the expression data of mature-miRNAs (rno-miR-9-5p, rno-miR-29a-3p, rno-miR-106a-5p, rno-miR-107 and rno-miR-125a-3p), which may be effective in AD, were taken with Real-Time PCR. KEY FINDINGS: Impairments occurred in behavioral tests of the rats in the O+D group. ßAP accumulation and AChE activity increased significantly in the forebrain in the O+D group compared to the C group. It was seen that Huperzine-A (Hup-A) reduced AChE activity and destructed ßAP accumulation. There was a significant decrease in the maximum contractile force at different frequencies in the O+D group and in the O group compared to the C group. SIGNIFICANCE: It was found that Hup-A contributed to the healing process in rats for damage occurring both in the brain and in the neuro-muscular system.