Disulfiram ameliorates bleomycin induced pulmonary inflammation and fibrosis in rats.

Bleomycin (BL) is a widely used anticancer drug that can cause pulmonary fibrosis due to increased fibroblast proliferation and increased secretion of extracellular matrix. RASSF1A is a tumor suppressor gene that is down-regulated by DNA methylation during fibrosis. Disulfiram (DSF), a noncytosine DNA methyltransferase inhibitor, can revert epigenetic biomarkers and re-express silenced genes. We investigated anti-inflammatory and anti-fibrotic effects of DSF on regulation of epigenetic molecules and histopathology in a rat model of BL induced pulmonary fibrosis. We used six groups of rats: sesame oil (SO) control (Co) group, BL group, BL + SO group and three BL + DSF groups administered 1 mg/kg DSF (BL + DSF), 10 mg/kg DSF (BL + DSF10) or 100 mg/kg DSF (BL + DSF100), respectively. BL was administered intratracheally to induce pulmonary fibrosis. DSF and SO were injected intraperitoneally (i.p.) 2 days before BL administration; these injections were continued for 3 weeks. At the end of the study, lung tissues were removed for molecular and histopathologic studies. Administration of 10 or 100 mg/kg DSF after BL induced pulmonary inflammation and fibrosis, and up-regulated RASSF1A and down-regulated TNF-α and IL-1 ß compared to the BL and BL + SO groups. A RASSF1A unmethylated band was observed using the methylation-specific PCR technique in rats that had been administered 10 and 100 mg/kg DSF, which indicated partial DNA demethylation. Histopathologic evaluation revealed that fibrosis and all inflammatory scores were decreased significantly in the BL + DSF10 and BL + DSF100 groups compared to the BL group. Our findings indicate that DSF modified DNA methylation by up-regulating RASSF1A, which reduced inflammation and fibrosis in BL induced pulmonary inflammation and fibrosis.

The efficacy of 2-formyl benzoic acid in reactivating diazinon inhibited murine cholinesterase.

Oximes, as classical acetylcholinesterase (AChE) reactivators, have some pharmacokinetics/pharmacodynamics disadvantages. During the synthesis of non-oxime compounds, we encountered the compound 2-formylbenzoic acid (2-FBA) with promising in vitro and in vivo cholinesterase (ChE) reactivating properties in the acute exposure to diazinon (DZN). For in vitro experiments, the healthy mice serum and brain homogenate were freshly prepared and exposed to DZN (160 µg/mL). After 10 minutes, 2-FBA was added to the poisoned samples, and ChE activity was measured afterward. For the in vivo assay, the mice were poisoned with DZN subcutaneous (SC) injection (50 mg/kg), and after 1 hour, either 2-FBA or Pralidoxime (2-PAM) was injected intravenously (IV). After 3 h, ChE activity was measured in the serum and brain homogenate samples. The LD50 (IV) for 2-FBA in mice was measured as well. 2-FBA effectively reactivated the inhibited ChE in serum and brain homogenate samples in vitro. In the in vivo experiments, while 2-FBA could significantly reactivate the brain ChE even better than 2-PAM, they failed to reactivate the serum ChE by single IV injection. LD50 of 2-FBA was calculated to be 963 mg/kg. There were no general toxicity signs in any treatment groups. The in silico results support the potential ability of 2-FBA efficacy via possibly Witting reaction mechanism. Our findings indicate that 2-FBA seems to be a suitable non-oxime candidate for AChE reactivation with minimal side effects. Further toxicokinetic studies on this compound are strongly recommended to be performed before conducting the clinical trial in humans.

Anti-Cancer Effect of Bromelain and Its Combination with Cisplatin on HN5 Cell Line (Squamous Cell Carcinoma).

Statement of the Problem: Squamous cell carcinoma (SCC) comprises over 90% of oral malignancies. Cisplatin, as a selective chemotherapy agent to treat SCC, has many side effects despite its high effectiveness. There are some studies on the effects of bromelain derived from pineapple stems on different malignancies. Purpose: The aim of this study was to investigate the effect of bromelain alone and in combination with Cisplatin on oral squamous cell carcinoma (OSCC) and fibroblast cell lines. Materials and Method: In this interventional study, the HN5 cell line of OSCC and fibroblast cell line were treated with different concentrations of bromelain alone and in combination with cisplatin. Cell viability test was performed after 24, 48 and 72 hours using MTT (3-)4,5-dimethylthiazol-2-yl(-2,5 diphenyl tetrazolium bromide) assay. In the final stage, the drug-treated cells underwent flow cytometry to assess apoptosis patterns. Data were analyzed using SPSS 17, ANOVA (for general comparison of groups) and LSD post hoc tests (for comparison two groups). p< 0.05 was considered statistically significant. Results: The findings suggested that although bromelain showed toxic effects on HN5 cancer cells, its combination with Cisplatin resulted in little improvement in its effectiveness. Bromelain alone and in combination with Cisplatin presented cytotoxic effects against fibroblasts, which depended on the dosage and time exposure (p< 0.05). The flow cytometry results did not support the superior effect of the combination of two medications over Cisplatin alone (p> 0.05). Conclusion: According to the findings, although adding bromelain to Cisplatin reduced toxicity on normal tissues, the combination of these two drugs did not increase the anticancer effect of Cisplatin. Thus, bromelain in combination with Cisplatin is not recommended as an adjuvant drug for OSCC.

Pre-Exposure to Radiofrequency Electromagnetic Fields and Induction of Radioadaptive Response in Rats Irradiated with High Doses of X-Rays.

Background: Some evidence shows that a pre-exposure to RF can mitigate the effects of subsequent exposures to high doses of ionizing radiation. Objective: We aimed to assess the effect of a pre-exposure to non-ionizing RF radiation on survival, weight changes, food consumption, and water intake of lethally irradiated rats. Material and Methods: In this case-control study, we used a commercial mobile phone (GSM, 900/1800 MHz) as well as a 2.4 GHz Wi-Fi router as the sources of pre-exposure to RF radiation. Forty-eight rats were randomly divided into six groups of control, "8 Gy X-rays", mobile phone, "mobile phone+8 Gy", Wi-Fi, and "Wi-Fi+8 Gy". Then, the survival fraction, weight loss, water, and food consumption changes were compared in different groups. Results: The survival analysis indicated that the survival rates in all of the exposed animals ("8 Gy X-rays", "mobile phone+8 Gy", "Wi-Fi+8 Gy") were significantly lower than the control, "Wi-Fi", and "mobile phone" groups. The changes in survival rates of "mobile+8 Gy", "Wi-Fi+8 Gy", and 8 Gy alone were not statistically significant. However, food and water intake were significantly affected by exposure to both RF pre-exposures and exposure to high dose ionizing radiation. Conclusion: To the best of our knowledge, the existence of a dose window for the induction of AR can be the cause of the lack of AR in our experiment. Our findings confirm that in a similar pattern with the adaptive responses induced by pre-exposure to ionizing radiation, the induction of adaptive response by RF-pre-exposures requires a minimum level of damage to trigger adaptive phenomena.

Recove® burn ointment for managing acute radiodermatitis in patients with breast cancer: A double blind randomized controlled trial.

Background: Radiodermatitis is the most common complication of radiotherapy. There is no gold standard for managing the radiodermatitis. This study aimed to evaluate the effect of topical Recove® burn ointment; basically compounded of sesame oil, camphor, and zinc oxide; in preventing acute radiodermatitis. Methods: This double blind RCT (IRCT No.: 201204047136N2) was performed on 71 patients that referred for radiotherapy after mastectomy to Shahid Rajaee Hospital (Babolsar-Iran) during 2013-2017. Patients were allocated into 2 groups; 34 in control group and 37 in Recove® group. Patients applied the ointment 2 times a day, before every radiation therapy session for 5 weeks. The radiation oncologist assessed the severity of dermatitis weekly for 5 weeks and graded it from 0 to 4 according to the RTOG criteria. Results: Baseline characteristics including age, and BMI had no significant difference between groups. The Recover group patients experienced significantly less severe dermatitis compared to the controls (p<0.001). None of the patients in Recove® group encountered more than grade 2 of RTOG criteria, however, in the control group, 4 (12.9%) patients experienced grade 3 of RTOG and 3 (9.7%) patients developed grade 4 of RTOG at the end of the 5th week. Conclusion: Our results indicate that Recove® ointment significantly reduces the severity of acute radiodermatitis.

Effects of arbutin on glucose uptake by glucose transporter 4 (GLUT4) and its cytoprotective properties in L6 skeletal muscle cell line.

It has been well known that oxidative stress and increased intracellular reactive oxygen species (ROS) have a pivotal role in disrupting the insulin signaling pathways leading to cellular insulin resistance. In this study, we evaluated arbutin's effects on glucose uptake by GLUT4 and cytoprotective properties in the L6 skeletal muscle cell line. The effect of arbutin and tertiary butyl hydrogen peroxide (t-BHP) on glucose uptake in cultured L6 cells was investigated by flow cytometry. We also evaluated gene expression levels of GLUT1 and GLUT4 in the L6 cells by quantitative real-time polymerase chain reaction analysis. The results from the study demonstrated that the optimum ROS generation occurred 3 h after 100 µM t-BHP treatment and pretreatment with arbutin (500 and 1000 µM) significantly inhibited the t-BHP induced ROS generation (p < .05). Our result indicated that 3 h pretreatment of L6 cells with 1000 µM of arbutin before 50 µM t-BHP significantly increased glucose uptake than the 50 µM t-BHP alone group (p < .05). Our findings may suggest that an increase in the uptake of 2-NBDG by L6 cells with arbutin pretreatment can be associated with increased expression of GLUT4 and GLUT1 under oxidative stress.

The efficacy of melatonin against radiotoxicity of iodine-131 and its response to treatment in hyperthyroid patients: a randomized controlled trial.

BACKGROUND: Since melatonin is a non-toxic compound with proven radioprotective effects, we aimed to investigate its efficacy in an in-vivo setting in hyperthyroid patients who are treated with iodine-131. This double-blind placebo-controlled study was conducted on hyperthyroid patients referred to nuclear medicine centers in Babol, Iran. We excluded patients suffering from hypertension treated with warfarin, autoimmune diseases, genetic diseases, cancers, smokers, chemical wounded, radiology and radiotherapy workers, and those who were treated with chemotherapy agents. Patients were randomly assigned to receive a capsule containing 300 mg of melatonin powder or a placebo. Just before receiving iodine-131, blood samples were taken from individuals. All 52 female patients received 10 to 20 mCi iodine-131 for treating hyperthyroidism. A second blood sample was taken one hour after the administration of iodine-131. MATERIAL AND METHODS: To determine the chromosomal damages before and after receiving radioiodine, we performed the cytokinesis- block micronucleus assay. Also, at phase 2, 6 months follow-up was performed, in which patients' positive responses to treatment were compared. RESULTS: The findings of this study indicate that the difference in micronucleus formation between the placebo and melatonin groups is not significant. However, a significant difference in the 6 months follow-up revealed that 61.5% and 85.7% of patients had a positive response to treatment in the placebo and melatonin groups, respectively. CONCLUSIONS: As one of the first studies dealing with the human in-vivo assessment on the radioprotective effects of melatonin, it was concluded that melatonin has a non-significant positive impact on reducing the rate of chromosomal damages in hyperthyroid patients treated with iodine-131. Nevertheless, the outcome of treatment was significantly higher by melatonin compared to the placebo group.

The effect of nonylphenol exposure on the stimulation of melanomacrophage centers, estrogen and testosterone level, and ERα gene expression in goldfish.

The present study tried to measure the formation of melanomacrophage centers (MMCs) in various organs of male and female goldfish exposed to nonylphenol (NP) and aimed to assess its relationship with the main sexual hormones, estrogen receptor expression, and the pigment content of the MMCs. Immature goldfish were exposed to 10-6 and 10-7 M NP for 25 days. After obtaining blood for measuring testosterone and estrogen (E2) levels, tissue samples were collected from various organs for histological studies, quantifying pigments using ImageJ software and chemical analysis, and measuring ERα gene expression. Results showed that the order of forming MMCs in various organs exposed to NP was liver > spleen > kidney, and the order of ERα gene expression was liver > testes > spleen > kidney in the male, and liver > spleen > kidney > ovaries in the female. Among the three pigments present in MMCs after exposure to the two doses of NP, melanin was more obvious (especially in the liver) and increased mostly in a dose-dependent manner in both sexes (especially in the male). Chemical analyses confirmed these results. Measurement of testosterone and E2 level in male and female goldfish showed that NP had more effect on the concentration of these hormones in male fish, indicating more endocrine-disrupting potential of NP against the male fish. Generally, the increase of melanin content of melanomacrophage centers coincided with the increase of ERα gene expression and decrease of testosterone level in goldfish after exposure to NP.

Evaluation of apoptotic effect of crocin, cisplatin, and their combination in human oral squamous cell carcinoma cell line HN5.

BACKGROUND: Squamous cell carcinoma (SCC) is the most common oral malignancy with high rate of mortality. Cisplatin, as the most effective chemotherapy drug, has side effects. Considering the studies on the use of crocin in saffron in the treatment of various malignancies, this study aimed at investigating the effects of crocin and cisplatin and their combination on SCC and fibroblast cell lines. MATERIALS AND METHODS: In this interventional study, HN5 and fibroblast cell lines were treated with different concentrations of crocin (12.5-50 µg/mL) and cisplatin (2, 4, 8, 16, and 32 µg/mL), and the cells were counted after 24, 48, and 72 h by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Data were analyzed with SPSS Version 17, and P < 0.05 was considered the level of significance. In the final stage, flow cytometry after 24 h in terms of the pattern of cell death was done. RESULTS: Both drugs had a toxic effect on malignant cells. One point was the high toxic effect of 8 µg/mL cisplatin not only on cancer cells (P < 0.001) but also on fibroblasts. However, combination with 12.5 µg/mL of crocin had the same effect on HN5 cell line, despite the less toxic effect in fibroblasts in comparison with cisplatin alone (P = 0.012). Apoptosis was the pattern of cell death showed by flow cytometry. CONCLUSION: Crocin in high concentrations can have not only significant toxicity in cancer cells but also side effects in healthy tissue. It seems that lower doses of crocin, in combination with cisplatin, besides having anticancer effect, can reduce the toxicity of cisplatin in healthy tissue.

Prenatal and breastfeeding exposure to low dose of diethylhexyl phthalate induces behavioral deficits and exacerbates oxidative stress in rat hippocampus.

Diethylhexyl phthalate (DEHP) is one of the most important derivatives of phthalate that has devastating effects on nervous system function. In this study, the effects of exposure with low doses of DEHP during pregnancy and lactation periods have been evaluated in rat's puppies. DEHP at doses 5, 40, 400 µg/kg/day and 300 mg/kg/day was given to mothers by gavage during pregnancy and lactation. The spatial and working memories were evaluated by Morris water maze test and Y maze, respectively. Oxidative stress levels were measured by biochemical tests. Histopathology of hippocampal tissue was assessed using hematoxylin and eosin, Nissl staining, and immunohistofluorescence in 60-days-old puppies. Behavioral data showed that low doses of DEHP decreased the working and spatial memories of male rats. Increased oxidative stress and decreased antioxidant activity were also observed in the hippocampus of rats which received the low doses of DEHP. However, neuronal damage, inflammation, and astrocyte activation were not significantly increased in the hippocampus of rats. Overall, exposure of mothers to low doses of DEHP during pregnancy and lactation cause behavioral deficits, especially in male newborn. The destructive effects of low doses of DEHP might be mediated through increased levels of oxidative stress in the brain.

Therapeutic effect of cold atmospheric plasma and its combination with radiation as a novel approach on inhibiting cervical cancer cell growth (HeLa cells).

Cervical cancer is one of the important cancers in women. Research on novel treatment approach can reduce the mortality and burden. Although radiotherapy is a common treatment, its negative side effects have concerned physician. In our study, we studied impact of cold atmospheric pressure plasma on the Hela cancer cells, as an alternative treatment. The effect of three different types of such plasma; dielectric barrier discharge (DBD), plasma jet, and afterglow plasma, on the cancer cells were studied. Moreover, some effective operating parameters such as exposure time, applied voltage, composition of working gas in plasma treatment were investigated on the survival of the afterglow plasma. Finally, treatments by the afterglow plasma, gamma radiation (1 Gy), and combination of both were compared. Analysis showed that DBD and plasma jet (direct exposure) effectively killed the cancer cells, even by a minimum applied voltage. But a fraction of the cells survived after the exposure of indirect diffused afterglow plasma. In the case of this plasma, we realized that higher applied voltage and exposure time led to less cell viability. Fewer fractions of survival cells were detected in the case of argon afterglow plasma comparing to oxygen afterglow. Cold atmospheric plasma and its combination with radiation therapy showed a significant decrease in viability of the cells, comparing to the radiation alone. Our research showed that plasma and its combination with radiation therapy have superiority over radiation therapy.

Quinazolinone-dihydropyrano[3,2-b]pyran hybrids as new α-glucosidase inhibitors: Design, synthesis, enzymatic inhibition, docking study and prediction of pharmacokinetic.

A series of new quinazolinone-dihydropyrano[3,2-b]pyran derivatives 10A-L were synthesized by simple chemical reactions and were investigated for inhibitory activities against α-glucosidase and α-amylase. New synthesized compounds showed high α-glucosidase inhibition effects in comparison to the standard drug acarbose and were inactive against α-amylase. Among them, the most potent compound was compound 10L (IC50 value = 40.1 ± 0.6 µM) with inhibitory activity around 18.75-fold more than acarboase (IC50 value = 750.0 ± 12.5 µM). This compound was a competitive inhibitor into α-glucosidase. Our obtained experimental results were confirmed by docking studies. Furthermore, the cytotoxicity of the most potent compounds 10L, 10G, and 10N against normal fibroblast cells and in silico druglikeness, ADME, and toxicity prediction of these compounds were also evaluated.

The Effect of Arbutin on The Expression of Tumor Suppressor P53, BAX/BCL-2 Ratio and Oxidative Stress Induced by Tert-Butyl Hydroperoxide in Fibroblast and LNcap Cell Lines.

OBJECTIVE: Arbutin (p-hydroxyphenyl-ß-D-glucopyranoside) possesses beneficial functions including antioxidant, antiinflammatory, and anti-tumoral activities. Due to the important role of oxidative stress and apoptosis in the successful treatment of cancer, understanding mechanisms that lead to apoptosis in cancer cells, is essential. The purpose of the current study was to evaluate the effect of arbutin on tert-butyl hydroperoxide (t-BHP)-induced oxidative stress and the related mechanisms in fibroblast and Lymph Node Carcinoma of the Prostate (LNCaP) cells. MATERIALS AND METHODS: In this experimental study, the LNCaP and fibroblast cell lines were pre-treated with arbutin (50, 250 and 1000 µM). After 24 hours, t-BHP (30 and 35 µM) was added to the cells. Viability was measured (at 24 and 48 hours) using MTT assay. The antioxidant effect of arbutin was measured by FRAP assay. The mRNA expression of P53 and BAX/BCL-2 ratio were measured using quantitative polymerase chain reaction (PCR). The percentage of apoptotic or necrotic cells was determined using a double staining annexin V fluorescein isothiocyanate (FITC) apoptosis detection kit. RESULTS: Arbutin pre-treatment increased the total antioxidative power and cell viability in the MTT assay and reduced BAX/BCL-2 ratio, P53 mRNA expression and necrosis in fibroblasts exposed to the oxidative agent (P<0.001). In addition, our results showed that arbutin can decrease cell viability, induce apoptosis and increase BAX/BCL-2 ratio in LNCaP cells at some specific concentrations (P<0.001). CONCLUSION: Arbutin as a potential functional ß-D-glucopyranoside has strong ability to selectively protect fibroblasts against t-BHP-induced cell damage and induce apoptosis in LNCaP cells.

Antibacterial and Cytotoxic Effects of Cyclodextrin-Triazole-Titanium Based Nanocomposite

Abstract In this paper, the antibacterial activity of triazole functionalized cyclodextrin (CD.Click) and cyclodextrin-triazole-titanium based nanocomposite (CD.COM) was evaluated. The results indicated that CD.Click and CD.COM perform a wide range of antibacterial activity against both gram positive (Staphylococcus aureus and Bacillus subtilis) and gram negative (Escherichia coli and Pseudomonas aeruginosa) bacteria. The cytotoxic effect of CD.COM was investigated in vitro on cancerous cell lines (cervical cancer, breast carcinoma and sarcoma osteogenic) and fibroblast cells by MTT assay. The cell viability evaluation confirmed that the growth of cancerous cells is inhibited in a dose and time dependent way without any significant effect on the normal fibroblast cells.

Combined Effects of Radiofrequency Electromagnetic Fields and X-Ray in Renal Tissue and Function.

PURPOSE: Biochemical and histopathological properties of renal tissues were reported to be affected by both radiofrequency electromagnetic fields (RF-EMF) and ionizing radiation. The radiation-induced changes in the kidney, including the serum levels of blood urea nitrogen (BUN) and creatinine (Cr), could lead to adverse health outcomes such as chronic kidney disease. These complications signify the importance of the research in this field. Thus, in this study, the effects of ionizing and non-ionizing radiations, as well as their combination, were assessed by evaluating the alteration in BUN, Cr, and histopathological changes in kidney tissue. MATERIALS AND METHODS: Ninety-six male Wistar rats were randomly divided into six groups and were exposed to either 900/1800MHz (mobile phone) or 2.4 GHz RF-EMF (Wi-Fi) radiation for 14 days, 8Gy x-ray, or their combination. Sera were collected from 2 mL of rat blood, then BUN and Cr levels were determined. Also, renal samples were stained with hematoxylin and eosin and evaluated histopathologically. RESULTS: Both BUN and Cr levels raised non-significantly after exposure to 8 Gy x-rays. Moreover, all measurements in the samples of x-ray groups were in borderline or higher than normal values. The BUN levels of control, Wi-Fi, x-ray, and Wi-Fi+x-ray groups were not significantly different. However, Cr levels in the Wi-Fi group were significantly higher than those of the controls, and BUN to Cr ratio levels were significantly lower than those of the controls. Also, tubular atrophy and vessel wall thickening were associated with these exposures. CONCLUSION: Exposure to 900/1800MHz, 2400 MHz EMF can alter the kidney function. However, pre-exposure to 900/1800MHz EMF could modulate the acute adverse effects of lethal x-ray dose, which addresses the adaptive response in the kidney.

Assessment of function, histopathological changes, and oxidative stress in liver tissue due to ionizing and non-ionizing radiations.

BACKGROUND: Compared to past decades, humans are exposed to rapidly increasing levels of radiofrequency electromagnetic radiations (RF-EMF). Despite numerous studies, the biological effects of human exposure to different levels of RF-EMF are not fully understood yet. This study aimed to evaluate the bioeffects of exposure to "900/1800 MHz" and "2.4 GHz" RF-EMFs, and x-rays alone as well as their potential interactions, i.e. inducing simple additive, adaptive, or synergistic effects. METHODS: 120 Wistar rats were randomly divided into ten groups of 12 each. The rats were exposed to RF-EMF, 10 cGy, and 8 Gy x-rays, a combination of these exposures, or only sham-exposed. The levels of liver enzymes were determined in serum samples by an auto-analyzer. Moreover, the histopathological changes, and the levels of malondialdehyde (MDA), nitric oxide, ferric reducing antioxidant power, total thiols, and protein carbonyl (PCO) were measured. RESULTS: Among the markers of liver function, gamma-glutamyltransferase was not associated with irradiation but, aspartate transaminase, alanine transaminase, and alkaline phosphatase showed some levels of association. MDA and PCO levels after 8 Gy irradiation increased, but pre-exposure to RF-EMF could modulate their changes. At the cellular level, the frequency of lobular inflammation was associated with the type of intervention. CONCLUSION: The exposure to both ionizing and non-ionizing radiations could alter some liver function tests. A short term pre-exposure to RF-EMF before exposure to an 8 Gy challenging dose of x-rays caused the alterations in oxidative stress markers and liver function tests, which indicate that oxidative stress is possibly involved in the adaptive response.

Decrease of intracellular ROS by arbutin is associated with apoptosis induction and downregulation of IL-1ß and TNF-α in LNCaP; prostate cancer.

Increased reactive oxygen species (ROS) along with inflammation are involved in the prostate cancer (PCa). Therefore, this study was conducted to investigate the molecular mechanisms that were affected by arbutin as an antioxidant on prostate cancer cell line; LNCap. The intracellular ROS measurement confirmed that arbutin significantly (p < .05) decreased the ROS levels in a dose-dependent manner. Detection of cell death profile established that 1,000 µM of arbutin could remarkably induced apoptosis (p < .05), while tert-butyl hydroperoxide (tBHP) as ROS inducer prompted necrosis. In addition, 1,000 µM of arbutin successfully decreased expressions of IL-1ß and TNF-α genes (p < .05). Furthermore, evaluation of the IL-1ß protein level showed that arbutin could significantly decrease this cytokine (p < .05). In summary, reduction of ROS along with increasing apoptosis and decreasing expression of pro-inflammatory genes following arbutin treatment can open new visions in the treatment of prostate cancer using complementary medicine. PRACTICAL APPLICATIONS: Nowadays, arbutin as a glycosylated hydroquinone is available commercially in both natural and synthetic forms. Arbutin is of interest because of its skin-lightening effect, and used in cosmetic products for cutaneous hyperpigmentation. Arbutin inhibited tyrosinase in melanocytes competitively. Moreover, arbutin was able to attenuate oxidative stress and, its anti-inflammatory activities has been established. In addition, arbutin has represented useful activities for suppression of malignant melanoma development. In addition, arbutin exhibits several pharmacological effects, including antimicrobial, antihyperlipidemic, antihyperglycemic, and alpha amylase inhibitory effects. In this study, we showed its effect on prostate cancer in vitro. Therefore, it opens new insights in the complementary medicine that can maintain or improve human health.

Systemic effects of starved fibroblast culture supernatant on immunosuppressed rats treated with cancer stem cells (LA7).

BACKGROUND: The present study aimed to investigate and compare the effect of starved fibroblast culture supernatant (SFS), DMEM and normal saline alone or along with LA7 on dexamethasone-treated immunosuppressed Wistar rats. METHODS: After the isolation of fibroblasts from the fresh foreskin of children, it was cultured in serum-free DMEM, and the supernatant collected after 16 hours (16h-SFS). This solution and the other treatments were injected subcutaneously into the rats from each group once daily for 14 days. The liver, intestine and lung histology along with blood cellular and biochemical characteristics were studied. RESULTS: The results showed that dexamethasone as immunosuppressant reduced the body weight. The histological change in the liver was mild fibrosis induced by LA7+16h-SFS. Also, among the different blood cellular and biochemical indices measured, the eosinophil percentage in the 16h-SFS treated rats , glucose levels in the 16h-SFS+LA7 group and triglyceride concentrations in the 16h-SFS group were changed (p<0.05). CONCLUSION: This study showed that the secretions of starved fibroblasts especially that combined with LA7 cancer stem cells could induce some minor histological and biochemical changes in immunosuppressed rats, and also it opened a new window for subsequent investigations on unknown mechanisms related to this work.

Targeting LA7 breast cancer stem cells of rat through repressing the genes of stemness-related transcription factors using three different biological fluids.

Down-regulation of stemness genes expression is important in differentiation therapy against cancer stem cells (CSCs). The aim of this study was to evaluate the Oct4 , Sox2, Nanog, and C-myc expression in rat breast cancer stem cells (LA7) which treated with human ovarian follicular fluid (FF), replicative senescent fibroblast culture supernatant (P14), and 16 h serum starved fibroblast supernatant (16 h-SFS). The cells were exposed to these biological fluids for 24 h, 72 h, and 7 days. Stem-loop RT-qPCR assay was used to quantify the expression of above mentioned genes. Results showed that FF had the least cytotoxic effect on the LA7 cells. Except for Nanog gene, exposure of LA7 cell line to 16 h-SFS and P14 decreased significantly expression of the three other genes after 24 h (P < 0.05). Nanog and Sox2 genes expression was also decreased in LA7 cells which have been already treated with FF for 24 h. Moreover, compared to the control solution, the expression of Oct4 increased significantly after 7 days exposure to FF (P < 0.05). Annexin V-PE /7-AAD-, acridine orange/ethidium bromide staining and doubling time assays revealed apoptosis and necrosis induction by these biological fluids in LA7 cells. Moreover, in an in vitro model of metastasis assay, i.e., scratch test, these fluids exhibited anti-LA7 migration activity which culminated in 16 h-SFS treated cells. Generally, this study showed that FF, 16 h-SFS, and P14 have positive effects on down-regulation of Nanog, Oct4, Sox2 and C-myc expression, and consequently can increase the differentiation of breast cancer stem cells. For the first time, this study provided some evidence indicating that some biological fluids have potential to differentiate the CSCs, show anti- survival, growth-, and cell migration activity.

A comparative study on immunophenotypic characterization and osteogenic differentiation of human mesenchymal stromal cells derived from periodontal ligament and gingiva.

BACKGROUND: Mesenchymal stem cells (MSCs) derived from periodontal ligament (PDL) and gingiva can be used for the development of cell-based regenerative approaches in dentistry and medicine. The purpose of this investigation was to establish a method for isolation of human stem cells from the PDL and gingiva, multilineage differentiation of those cells, and comparison of periodontal ligament mesenchymal stem cells (PDLMSCs) and gingival mesenchymal stem cells (GMSCs). METHODS: PDL and gingival tissues of third molar teeth were digested enzymatically and the proliferative potential of human PDLMSCs and GMSCs was compared by MTT assay. The expression of cell surface epitopes was analyzed by flow cytometry. To investigate the multilineage differentiation capacity of these stem cells, osteogenic and adipogenic differentiation was achieved. The specific staining of nodules was performed to evaluate differentiation, whereas the expression of alkalin phosphatase (ALP) and collagen A I (COL I) genes was analyzed by quantitative real-time polymerase chain reaction. RESULTS: The outgrown cells derived from PDL and gingival tissues were similar, fibroblast-like, and spindle-shaped. Further, the proliferation potential of GMSCs was greater than PDLMSCs. Both types of stem cells expressed MSC precursor markers, including CD73, CD90, and CD105, whereas they were negative for hematopoietic markers, including CD34 and CD45. PDLMSCs demonstrated more osteogenic potential compared to GMSCs with strong mineral noduls, and significantly greater expression of up-regulated bone-related markers ALP and COL I. CONCLUSION: MSCs derived from PDL and gingiva demonstrated multipotent characteristics, suggesting new therapeutic approaches in tissue engineering and PDLMSCs are more appropriate candidates for this purpose.