Effects of Escherichia coli strain Nissle 1917 on arsenic-challenged goldfish (Carassius auratus): histological evidence.

Arsenic (As) contamination in natural water resources has become a great disaster throughout the world posing serious health problems. The current study was performed to evaluate the protective effects of Escherichia coli strain Nissle 1917 (EcN) against As exposure in goldfish (Carassius auratus). Fish were fed three times a day with 4.00% of body weight of diet with different doses (0.00, 1.00 × 106, 1.00 × 107 and 1.00 × 108 CFU g-1) of EcN for 80 days and then, challenged with 20.00 mg L-1 As for 96 hr under stagnant flow. Physicochemical characteristics of the inlet water were temperature of 25.10 ± 0.70 ˚C, pH of 7.30 ± 0.20 and dissolved oxygen of 7.30 ± 0.30 mg L-1 and 50.00% of water was exchanged once a week. Afterwards, fish were euthanized with a clove oil solution (50.00 µL L-1) and tissues were dissected from each fish and immediately fixed in 10.00% buffered formalin. The histopathological results indicated that the supplemented EcN did not have any side effects on various organs. It was also observed that the damages to kidney, liver, gill and skin were pronounced in fish exposed to As. However, the histopathological damages induced by As in fish tissues were less pronounced in the EcN-treated groups compared to the fish fed with the basal diet. Lamellar blood congestion in gills and epidermal cells detachment from the skin surface as well as hepatocytes, enterocytes and tubular necrosis were reduced in treated groups. These findings indicate that EcN has the potential to ameliorate the As-induced organ toxicity.

Dietary astaxanthin (Lucantin® Pink) mitigated oxidative stress induced by diazinon in rainbow trout (Oncorhynchus mykiss).

The potential of commercial astaxanthin on growth, biochemical factors, and antioxidant-related gene expression following a challenge with diazinon were studied in rainbow trout (Oncorhynchus mykiss). Fish (~ 20.70 g) were fed diets containing commercial astaxanthin (ASX) at 0.00 (CTR and ASX0), 0.50 (ASX1), 2.00 (ASX2), and 5.00 (ASX3) g kg-1 for 60 days. Afterwards, the treated fish (ASX1, ASX2, ASX3) as well as the fish in ASX0 group were challenged with diazinon (0.11 mg L-1) for 96 hr whereas fish in the CTR group was not challenged with diazinon. Results showed that growth pattern improved significantly with all enriched diets compared to the ASX0 group. Metabolic enzyme activities, including alanine aminotransferase and alkaline phosphatase decreased in ASX2 and ASX3 groups with respect to the ASX0 group. Serum antioxidant status also showed the same pattern with enhancement in the fish fed with the ASX2 and ASX3 supplemented diets. Feeding the fish with astaxanthin, particularly in the ASX3 group, up-regulated the expression of some antioxidant-relevant genes, including superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione S-transferase (GST), and nuclear factor erythroid-2 related factor 2 (Nrf2) in the kidney and liver. Besides, the histopathological damages in kidneys and liver induced by diazinon were less pronounced in the ASX2 and ASX3 groups compared to the ASX0 group. In conclusion, commercial astaxanthin, especially at 5.00 g kg-1, enhanced the growth performance and ameliorated the oxidative stress induced by diazinon in rainbow trout.

The effect of co-administration of artemisinin and N-acetyl cysteine on antioxidant status, spermatological parameters and histopathology of testis in adult male mice.

OBJECTIVES: This in vivo study aimed to evaluate the effect of various concentrations of artemisinin (Art) alone or together with N-acetyl cysteine (NAC) on spermatological indices, antioxidant status, and histopathological parameters of testicular tissue in adult male mice. METHODS: Six groups of five healthy male mice (25-30 g) were randomly assigned to different experimental groups. These groups received DMSO and corn oil (0.1%) as an Art solvent (Control), 50 mg kg-1 Art (Art-50), 250 mg kg-1 Art (Art-250), 50 mg kg-1 Art + 150 mg kg-1 NAC (Art-50+NAC-150), 250 mg kg-1 Art + 150 mg kg-1 NAC (Art-250+NAC-150) and 150 mg kg-1 NAC (NAC-150) for a period of 7 days. Testes and epididymis were prepared to evaluate the malondialdehyde (MDA), glutathione (GSH), superoxide dismutase (SOD), catalase (CAT), peroxidase (POX), spermatological indices, and histological parameters. RESULTS: We showed that the high dose of Art (Art-250) significantly reduced the sperm count, motility, viability, and the activity of CAT and increased the levels of MDA compared to the control group. Also, the overdose of Art caused adverse changes in testicular tissue. Co-administration of NAC with Art (Art-250+NAC-150) corrected the adverse effects of Art. CONCLUSIONS: The current study reports that a high dose of Art affects, spermatological parameters, antioxidant/stress oxidative status of the male reproductive system, and NAC is capable neutralize all adverse effects caused by Art.

In Vivo Toxicity of Oral Administrated Nano-SiO2: Can Food Additives Increase Apoptosis?

Nano-silicon dioxide (nano-SiO2) has a great deal of application in food packaging, as antibacterial food additives, and in drug delivery systems but this nanoparticle, despite its wide range of utilizations, can generate destructive effects on organs such as the liver, kidney, and lungs. This study is aimed at investigating the toxicological effects of nano-SiO2 through apoptotic factors. For this purpose, 40 female rats in 4 groups (n = 10) received 300, 600, and 900 mg/kg/day of nano-SiO2 at 20-30 nm size orally for 20 days. Relative expression of Caspase3, Bcl-2, and BAX genes in kidney and liver was evaluated in real time-PCR. The results indicated the overexpression of BAX and Caspase3 genes in the liver and kidney in groups receiving 300 and 900 mg/kg/day of nano-SiO2. Bcl-2 gene was up-regulated in the liver and kidney at 600 mg/kg/day compared to the control group. Overexpression of the Bcl-2 gene in the kidney in 300 and 900 mg/kg/day recipient groups was observed (P ≤ 0.05). Histopathological examination demonstrated 600 mg/kg/day hyperemia in the kidney and lungs. In addition, at 900 mg/kg/day were distinguished scattered necrosis and hyperemia in the liver. The rate of epithelialization in the lungs increased. The nano-SiO2 at 300 and 900 mg/kg/day can induce more cytotoxicity in the liver and lung after oral exposure. However, cytotoxicity of nano-SiO2 at 600 mg/kg/day in the kidney and lung was noticed. Hence, the using of nano-SiO2 as an additive and food packaging should be more considered due to their deleterious effects.

Trichoderma harzianum as fungicide and symbiont: is it safe for human and animals?

Trichoderma species are considered as biological control agents against numerous phytopathogenic fungi. They are also helpful for plants as plant symbiont. This study aimed to identify harmful effects of Trichoderma in laboratory animals. In the first step, inhalation toxicity was studied. Six rats as control received a spray of bio-formulation without spores. Ten rats as treatment A received    1.00 × 106 colony-forming unit (CFU) of Trichoderma spores and ten rats as treatment B received 1.00 × 107 CFU per test of Trichoderma spores. The harmful effects of Trichoderma were obvious especially in the lungs, liver and kidney, and some blood parameters were abnormal. In the second step, we studied acute oral toxicity by gavage. Four rats as control received bio-formulation without spores. Six rats as treatment A received 1.00 × 106 CFU per test of Trichoderma spores. Six rats as treatment B received 1.00 × 107 CFU per test of Trichoderma spores. The harmful effects of Trichoderma were noticeable more in the liver and kidney tissues. For dermal toxicity study, two rabbits as control received bio-formulation without spores by rubbing on the surface of the skin. Treatment groups A and B received 1.00 × 106 and 1.00 × 107 CFU per test of Trichoderma spores, respectively (four rabbits for each group). The liver and kidney and some blood parameters were abnormal. Trichoderma has some harmful effects on tissues and organs and although it is a natural product, it should be used under cautions.

The effects of L-carnitine-loaded solid lipid nanoparticles on performance, antioxidant parameters, and expression of genes associated with cholesterol metabolism in laying hens.

The purpose of this study was to investigate the production performance, antioxidant parameters, egg yolk cholesterol content, and expression of genes related to cholesterol metabolism in laying hens fed L-carnitine (LC) and L-carnitine-loaded solid lipid nanoparticles (LC-SLNs). A total of 350 Hy-Line (w-36) laying hens at 50 wk of age (1520.0 ± 0.7 g) were randomly assigned to 35 units (5 replicates and 50 hens in each treatment) with seven dietary treatments as a completely randomized design. The dietary treatments were corn-soybean meal-based diets, including 1) Control (basal diet); 2) Basal diet +50 mg/kg LC (50LC); 3) Basal diet +100 mg/kg LC (100LC); 4) Basal diet +150 mg/kg LC (150LC); 5) Basal diet +50 mg/kg LC-SLNs (50LC-SLNs); 6) Basal diet +100 mg/kg LC-SLNs (100LC-SLNs) and 7) Basal diet +150 mg/kg LC-SLNs (150LC-SLNs). Results showed that the 50LC-SLNs had the least feed conversion ratio (FCR) in all groups (P < 0.05). The dietary supplementation of 100LC-SLNs decreased (P < 0.01) the egg yolk cholesterol concentration from 14.71 to 11.76 mg/g yolk (25%). The 50LC-SLNs group produced the most total antioxidant capacity with a difference of 58.44% compared to the control group (P < 0.01). The greatest amount of total superoxide dismutase was found for 50LC-SLNs (P < 0.05), while the glutathione peroxidase was not affected by the experimental treatments (P > 0.05). Serum malondialdehyde levels were reduced by 50.52% in laying hens fed 50LC-SLNs compared to the control group (P < 0.05). The transcript level of 3-hydroxy-3-methylglutaryl coenzyme A reductase was significantly decreased (P < 0.01) in the LC and LC-SLNs groups. The expression of cholesterol 7α-hydroxylase was significantly increased (P < 0.01) in the plain LC (∼83%) and LC-SLNs (∼91%) groups. The inclusion of LC-SLNs in the diet increased (P < 0.05) the villus height and decreased villus width in all three parts of the small intestine. Dietary inclusion of LC was found to reduce egg yolk and serum cholesterol content by improving the production performance and antioxidant status. The LC-SLNs groups were more affected than the plain LC groups, which may be attributed to the increased bioavailability of LC.

The Protective Effect of Artemisia spicigera Ethanolic Extract against Cryptosporidium parvum Infection in Immunosuppressed Mice.

BACKGROUND: A specific treatment has not yet developed for cryptosporidiosis, and some of the used drugs had side effects in immunodeficient patients. The goal of an appropriate remedy is to remove symptoms and improve immune responses in hosts. The current study was designed to evaluate the therapeutic efficacy of Artemisia spicigera ethanolic extract in experimentally infected immunosuppressed mice. METHODS: Thirty six NMRI mice, 4-6 wk old, were randomly divided into six equal groups. C1: uninfected, treated control; C2: infected, untreated control; T1, T2, T3, and P: infected, treated with 0.2, 2, and 20 mg/ml extract, and 5mg/ml paromomycin, respectively. Mice were experimentally infected by oral administration of 104 oocysts/animal of Cryptosporidium parvum and treated orally for eight days per 12h, starting 12h before experimental infection. The presence of oocyst shedding, weight gain/loss, and the histopathology of ileum sections were examined. RESULTS: Results revealed that oocyst shedding was significantly (P<0.05) reduced in treatment groups. There was no significant difference between the mean of weight gain/loss in the infected control and treated groups. Histopathological analysis of ileum sections further supported the parasitological findings. CONCLUSION: Artemisia spicigera had acceptable efficacy as a therapeutic agent for cryptosporidiosis.

The Roles of Lactobacillus Acidophilus and Pectin in Preventing Postoperative Sepsis and Intestinal Adaptation in a Rat Model of Short Bowel Syndrome.

One of the primary causes of morbidity and mortality in those with short bowel syndrome (SBS) is sepsis, caused by bacterial translocation (BT). Since synbiotics can cease gut-related bacterial overgrowth, they may serve as a supportive dietary supplement-based strategy after gastrointestinal surgery. This study was conducted to determine the effects of Lactobacillus acidophilus and pectin on BT and gut adaptation after extensive small bowel resection in the rat. Forty rats were distributed in four groups. Group A suffered laparotomy, group B suffered gut transection and reanastomosis, SBS rats (group C) suffered 75% small gut resection, and finally, Group D suffered gut resection and treated with a synbiotic cocktail from day 7 before the surgery to day 14 after it. Intestinal structural changes and BT to mesenteric lymph nodes, liver, portal blood, and peripheral blood were detected on day 15 post-surgery. Treatment with a synbiotic cocktail led to a considerable reduction in bacterial translocation to liver and portal vein (degree II) compared with SBS untreated rats. Also, synbiotic administration significantly increased jejunum and ileum villus height and crypt depth, ileum villus width, and percentage of goblet cells in jejunum and ileum compared with SBS rats. In the rat model of short bowel syndrome, L. acidophilus, and pectin, as a potential synbiotic compound, could decrease the BT from the gut and improve the bowel adaptation.

Differential expression of small heat shock proteins in the brain of broiler embryo; the effects of embryonic thermal manipulation.

Broilers are more vulnerable to high temperatures than mammals due to the feather cover, lack of sweat glands, fast growth and intensive breeding in commercial systems. Thermal stresses affect the function of various organs and change the expression profiles of hundreds of genes in the different tissues of broilers. Thermal manipulation (TM) during embryogenesis can increase heat tolerance in growing broilers. Small heat shock proteins (SHSPs) are a group of HSPs which participate in many cellular functions like response to different stressors. However, their role in the thermotolerance has not been fully elucidated. Ninety fertilized eggs were randomly divided into three groups (30 eggs/group; 10 eggs/replicate). Normal control (NC) eggs were incubated at 37.5 °C throughout the incubation period whereas heat stress (HS) and cold stress (CS) groups were kept at 41 °C and 33 °C from 15 to 17th day of incubation for 3 h each day, respectively. On day 20, samples from the cerebrums were harvested for histopathology and mRNA expression analyses of HSPB1, HSPB5, HSPB8, and HSPB9. There were no significant differences in survivability, defected embryos, hatchability, and body weight among treatments. TM had no major deleterious effects on the cerebral tissue except for mild degeneration in the HS group. HSPB1, HSPB5, HSPB8, and HSPB9 were expressed in the presence and absence of TM. All SHSP genes tested were downregulated in response to TM except for HSPB9 which was upregulated in the HS group. The highest change in gene expression due to TM observed for HSPB1. This study presents a broader understanding of mechanisms underlying response to TM in broilers. The results suggest that HSPB1, HSPB5, HSPB8, and HSPB9 are involved in thermotolerance in broilers and SHSPs could be involved in the gene expression profiling of TM. It may propose the use of nutritional supplements in the poultry industry to modulate SHSPs.

Pathological characteristics of Linguatula serrata (aberrant arthropod) infestation in sheep and factors associated with prevalence in Iran.

Linguatula serrata is a cosmopolitan zoonotic parasite in which carnivores and herbivores serve as final and intermediate hosts, respectively. The aim of this study was to compare the L. serrata nymphal infection rate and intensity of infection (mean number of nymphs ±â€¯standard error) to the appearance and pathological changes of mesenteric lymph nodes (MLNs) infected with L. serrata using sheep slaughtered in Tabriz, Iran. In addition, the effect of age, sex, and season on the prevalence of L. serrata infection was evaluated. For this purpose, over a four-year period, 31,078 MLNs from 3199 sheep were examined, with 4972 (15.99%) MLNs infected representing 518 (16.20%) sheep. Collected MLNs were categorized by color as normal, red or black and by consistency as normal, soft or hard. L. serrata were found in 8.88% of normal-colored MLNs, 14.45% of red (hemorrhagic) MLNs and 44.57% of black-colored MLNs, with the difference being significant for infection and infection intensity (P < 0.0001). In regards to MLN consistency, 7.98% of normal, 31.52% of soft and 5.42% of hard lymph nodes were found to be infected with the infection rate and intensity in soft nodes being significantly different (P < 0.0001). Pathological changes in MLNs infected with L. serrata nymph with normal color and consistency had calcification of the L. serrata nymph, granulomatous inflammation around the nymph and some neutrophils. Granulomatous inflammation around the L. serrata nymph, haemosiderophage, macrophage and lymph node depletion from lymphocytes were observed in MLNs infected with L. serrata nymph with soft consistency. In addition, MLNs infected with L. serrata nymph with hard consistency and black color contained neutrophils in the capsule's wall, caseous necrotic mass and L. serrata surrounded by a thick capsule. In regards to prevalence, age, sex and season (autumn) were significant (P < 0.0001, P < 0.01, respectively). These study results suggest that targeted meat inspection and targeted animal interventions could be used to decrease human exposure to L. serrata and animal infection.

Curcumin and Nanocurcumin Oral Supplementation Improve Muscle Healing in a Rat Model of Surgical Muscle Laceration.

OBJECTIVE: To compare the effects of curcumin and nanocurcumin oral supplementation on the muscle healing rate of an animal model of surgical muscle laceration. METHODS: Thirty-two male adult rats were randomly divided into sham, control, curcumin, and nanocurcumin groups. Partial transection of the gastrocnemius muscle was made in the right limb of the control and treatment groups. The sham and control groups received normal saline, curcumin group received 500 mg/kg of curcumin and nanocurcumin group received 100 mg curcumin-loaded nanomicelles orally every day. They euthanized two weeks later and the specimens were stained by hematoxylin-eosin (H&E) and Masson's trichrome methods. Aspartate transaminase (AST) and creatine phosphokinase (CPK) were measured in blood samples. RESULTS: The percentage of collagen fibers in the nanocurcumin group was significantly lesser than the control and curcumin groups (p<0.001). Muscle fiber regeneration in the treatment groups was significantly higher than the control group (p<0.001). The blood vessels of the nanocurcumin group were significantly more than other groups (p<0.001). Plasma AST had a significant difference in the control group compared to the sham and nanocurcumin groups (p=0.026). The plasma CPK level of the control group was also significantly higher than other groups (p<0.001). CONCLUSION: In conclusion, although oral curcumin supplementation has little effects because of its poor bioavailability, embedding it in nanoparticles could enhance its systemic effects in promoting the muscle healing process.

Effects of L-carnitine and betamethasone on ischemia-reperfusion injuries and sperm parameters following testicular torsion in a rat model.

Testicular torsion is a consequence of spermatic cord twisting which causes progressive damage to the structure of the testis and reduces sperm quality and usually results in infertility. In the present study, with the assumption of the protective effects of L-carnitine and betamethasone against ischemia-reperfusion (IR) injuries, their effects on twisted testicles were evaluated and compared. Twenty Wistar rats were randomly divided into four groups and used in this study. Except for the Sham (S) group, testicular IR was induced surgically in three other groups, including Control (C), Betamethasone (BM), and L-carnitine (LC) groups. Betamethasone and L-carnitine were injected before detorsion in the BM and LC groups, respectively. After twelve hours of reperfusion, the testicles were detached, and prepared for sperm parameters evaluation such as sperm count, motility, viability, morphology, and chromatin quality, and histopathologic evaluations, including mean seminiferous tubular diameter (MSTD), germinal epithelial cell thickness (GECT), and Johnsen's mean testicular biopsy scoring (MTBS). The MSTD, GECT, and healthy sperms in the C group were significantly lower than the other groups, while the BM and LC groups were significantly different from others in MTBS. The number of sperms and sperm motility in the BM group was significantly higher than the C group. Sperm viability in the BM and LC groups were significantly higher than the C group. The results of this study showed that both L-carnitine and betamethasone similarly can be effective in treating testicular IR injuries.

The effects of coadministration of tilorone dihydrochloride and culture supernatants from Lactobacillus reuteri on the mouse hepatoma cell line.

CONTEXT: Tilorone dihydrochloride is a therapeutic agent with a different mechanism in cancer. The species of Lactobacillus have an important role in cytotoxic effect. AIMS: Because of unknown effects of tilorone and culture supernatants from Lactobacillus reuteri on hepatoma, the aim of this study is to evaluate apoptotic, cytotoxic, and therapeutic effects of tilorone on mouse hepatoma cell line with and without culture supernatants from L. reuteri. MATERIALS AND METHODS: To do so, after cell line culture, cells were divided into different groups such as negative control, treatment with four doses of tilorone, positive control of supernatant (single dose), and combination therapy groups of different doses of tilorone with supernatant (constant doses), for 48 h. All groups were studied with pathologic tests, biochemical study, tetrazolium dye (3-(4, 5- dimethylthiazol -2-yl)-2, 5-diphenyltetrazolium bromide [MTT]) assay, and absolute real-time-polymerase chain reaction (RT-PCR) were done to assess Bax and Bcl-2 genes expression, as molecular studies. RESULTS: MTT assay results revealed that the tilorone tissue culture IC50 (TCIC50) on the Hepa1-6 cell line was 50 µg/ml. RT-PCR analysis showed that tilorone dihydrochloride induced upregulation and downregulation in expression of Bax and Bcl-2, respectively. Simultaneous, antioxidant effect has also seen in a way that prevented necrosis, in biochemical analysis. These results were dose dependent and statistically significant compared to the control group. CONCLUSIONS: Based on these results, it appeared that this agent could be a good candidate for further evaluation as effective chemotherapy acting through the induction of apoptosis in hepatoma. The cell death caused through bacterial supernatant was rather necrosis than apoptosis.

Beneficial effects of killed Tsukamurella inchonensis on rainbow trout (Oncorhynchus mykiss) growth, intestinal histology, immunological, and biochemical parameters.

Present study was conducted to investigate the effects of heat-killed Tsukamurella inchonensis on growth performance, gastrointestinal structure, immune response, and biochemical parameters in rainbow trout. Fish (mean weight 25 g) were fed basal diet (control), diets containing 2.48 × 108 colony-forming units (low-dose group) or 1.24 × 109 colony-forming units (high-dose group) of heat-killed Tsukamurella inchonensis per 1 kg of feed for 90 days. Results showed that growth performance was significantly enhanced in both treatment groups compared to the control group. The intestinal villus length and pyloric cecal fold length were mainly enhanced in the high-dose group. On the other hand, higher goblet cell percentage was shown with administration of dead Tsukamurella inchonensis in both treatment groups. Immune parameters such as alternative complement activity, immunoglobulin level, and hemagglutination titer were significantly higher in treatment groups than in fish fed in the control diet. Meanwhile, feeding heat-killed Tsukamurella inchonensis especially at higher dose caused a decrease in the levels of total cholesterol, uric acid, and lipid peroxidation product whereas no significant changes were noted in serum-specific marker enzymes levels, namely alanine aminotransferase (ALT), alkaline phosphatase (ALP), and aspartate aminotransferase (AST) by feeding both treatment diets compared to the control group. This study suggests that heat-killed Tsukamurella inchonensis especially at 1.24 × 109 colony-forming units had more potential to enhance growth, immunological parameters, and intestinal structure in rainbow trout.

Bone marrow mesenchymal stem cells modified pathological changes and immunological responses in ovalbumin-induced asthmatic rats possibly by the modulation of miRNA155 and miRNA133.

In the current experiment, we investigated the immune-modulatory potential of mesenchymal stem cells (MSCs) and conditioned media (CM) in attenuating of chronic asthmatic changes in a rat model. Male rats were divided into control (C) and ovalbumin-sensitized (S) groups, which further allocated into three subgroups; rats received systemically 50 µl volume of PBS (C and S groups), CM (CSV and SSV groups) and rats received intravenous infusion of 2 × 106 bone marrow-derived mesenchymal stem cells (rBMMSCs) (CCV and SCV groups). Two weeks later, the expression of interleukin (IL)-4, IL-13, and IL-10, miRNA133, and miRNA155 was measured by real-time PCR. Pathological changes and the recruitment of rBMMSCs into pulmonary parenchyma were evaluated by histopathological and immunofluorescence analyses, respectively. The systemic injection of rBMMSCs, but not CM, decreased the levels of IL-4, IL-13, IL-10, miRNA133, miRNA155 and reduced pathological changes in sensitized rats as compared with other sensitized groups (p < 0.001 to p < 0.05). rBMMSCs transmigrated to lung tissue in cell-administrated rats, albeit intensity of asthmatic changes, in turn, affected the amount of recruited cells. Collectively, our data suggest the potential role of MSCs, but not CM, in reducing pathological changes possibly via the modulation of miRNA133 and miRNA155 during asthmatic changes.

Effects of coadministration of artemisinin and iron on histopathologicalalterations in the AGS gastric adenocarcinoma cell line.

BACKGROUND/AIM: The aim of the present study was to examine the anticancer effects of combination treatment with artemisinin and iron on the human gastric cancer cell line. MATERIALS AND METHODS: The AGS cell line was cultivated separately as a monolayer in culture flasks and different doses of 99% pure artemisinin with invariable doses of iron sulfate were added to the culture media and adhesive cells on the flask's bottom were stained with hematoxylin-eosin for a pathological assay. RESULTS: Damage to cancerous cells increased dose dependently and it was higher in the combination treatment groups (artemisinin plus iron). The histopathological changes were observed specially in the groups with high artemisinin concentration as cell swelling, nucleus swelling, and formation of small and large vacuolization. Necrotic changes as nucleus pyknosis were seen too. Changes in groups receiving both artemisinin and iron gradually became more severe with dose increase. CONCLUSION: Pathologic studies showed that the cytotoxic effects of artemisinin were dose dependent and the presence of iron enhanced the artemisinin's anticancer potency.

Bone marrow mesenchymal stem cells and their conditioned media could potentially ameliorate ovalbumin-induced asthmatic changes.

BACKGROUND: The major feature of asthma is governed by chronic airway inflammation. This investigation was proposed to achieve the suitable candidate for ameliorating long-term chronic asthmatic changes of respiratory tract. METHODS: 36 rats were classified into healthy (C) and ovalbumin (OVA)-sensitized animals (S). To sensitize, the rats were exposed to OVA over a course of 32±1days. One day after sensitization, equal six different groups were subjected to experimental procedure (n=6); Rats only received intratracheally 50ml PBS (CPT and SPT groups), 50µl conditioned medium (CM) (CST and SST groups) and 50µl PBS containing 2×106 rat bone marrow-derived mesenchymal stem cells (rBMMSCs) (CCT and SCT groups). Two weeks after treatment, tracheal responsiveness, immunologic responses and recruitment of rBMMSCs into the lung as well as pathological changes were evaluated. RESULTS: A high degree of tracheal responsiveness, total white blood cell and percentages of eosinophil and neutrophil was significantly recorded in all sensitized groups rather than of controls (p<0.001 to p<0.05). Of interest, all above-mentioned parameters decreased significantly in SST and notably SCT groups as compared to S group (p<0.001 to p<0.05). The results revealed decrease number of blood CD3+CD4+ and concurrent increase in CD3+CD8+ in all sensitized rats as compared to control (p<0.001 to p<0.05). Noticeably, no significant modulatory effects of either cell or CM administration were achieved on the CD3+CD4+ and CD3+CD8+ populations in non-asthmatic rats. Moreover, the number of CD3+CD4+ in SST and SCT groups tended to increase, which coincided with a decreased manner of CD3+CD8+ populations as compared with S group (p<0.001 to p<0.05). However, the CD3+CD4+ cells in SCT rats were significantly higher than the group SST (p<0.01) whereas CD3+CD8+ cells diminished simultaneously (p<0.001). Real-time PCR analysis further showed that both CM and particularly MSCs changed the expression of interleukin (IL)-4 and IL-10 in the asthmatic groups to the near level of control rats (p<0.001 to p<0.05). Histopathological analysis revealed a profound reduction of lungs injuries in asthmatic rats when received CM and peculiarly mesenchymal stem cells (p<0.01 to p<0.05). CONCLUSION: Our study shed light on the superior effects of rBMMSCs, rather than CM, in attenuating of chronic asthmatic changes in the rat model.

Irradiation enhances susceptibility of tumor cells to the antitumor effects of TNF-α activated adipose derived mesenchymal stem cells in breast cancer model.

Gene modified or cytokine activated mesenchymal stem cells (MSCs) have been used as a treatment in various types of cancer. Moreover, irradiation is usually applied as either a standard primary or adjuvant therapy. Here, we showed that the expression of TNF related apoptosis-inducing ligand (TRAIL) and Dickouf-3 (Dkk-3), the promising anticancer proteins, increased in murine adipose-derived mesenchymal stromal cells (AD-MSCs) following activation with TNF-α, resulting in the induction of apoptosis in cancer cells. Also, anticancer effects of TNF-α activated AD-MSCs were intensified with irradiation. In vivo results showed that TNF-α preactivated AD-MSCs combined with irradiation decreased tumor size and increased survival rate in tumor bearing mice. On the other hands, both TNF-α preactivated AD-MSCs with or without irradiation prevented metastasis in ling and liver, and increased apoptosis in tumor mass. Finally, flowcytometry assay demonstrated that naïve AD-MSCs combined with irradiation but not TNF-α activated MSCs with irradiation increased Treg population in lymph node and spleen. Altogether, obtained results suggest that TNF-α activated MSCs combined with irradiation therapy can serve as new strategy in breast cancer therapy.

The Protective Effect of α-Hederin, the Active Constituent of Nigella sativa, on Lung Inflammation and Blood Cytokines in Ovalbumin Sensitized Guinea Pigs.

In the present study, the preventive effect of two different concentrations of α-hederin, the active constituent of Nigella sativa, on lung inflammation and blood cytokines in ovalbumin sensitized guinea pigs was examined. Forty eight male adult guinea pigs were divided into control (C), sensitized (S) and sensitized pretreated groups; with thymoquinone (S+TQ), low dose (S+LAH) and high dose of α-hederin (S+HAH) and inhaled fluticasone propionate (S+FP). The lung histopathology and blood levels of IL-4, IFN-γ and IL-17 were assessed. Compared to sensitized animals, all pathological changes improved significantly in pretreated groups (p < 0.001 to p < 0.05). These improvements in α-hederin pretreated groups were similar to S+TQ and S+FP groups except cellular infiltration in S+LAH and S+HAH groups which was lower than S+TQ group (p < 0.05). The blood IL-4 and IL-17 levels in S+HAH groups showed a significant decrease compared to S group (p < 0.05) which were similar to S+TQ and S+FP groups. The level of IFN-γ in S+LAH and S+HAH groups increased significantly compared to S group (p < 0.05) which was higher than S+FP group (p < 0.05). Blood IL-4 in S+HAH group was significantly lower than S+LAH group (p < 0.05). In conclusion, α-hederin could attenuate the lung inflammation and improve the changes of cytokines like thymoquinone and fluticasone in used dosages.