Melatonin Protects Against Titanium Oxide-Induced Neurotoxicity: Neurochemical, Neurobehavioral, and Histopathological Evidences.

titania (titanium dioxide, TiO2) is known to induce neurotoxicity and CNS dysfunctions. Numerous studies have explored the neuroprotective effects of melatonin against neurotoxicity. This study evaluates the potential of melatonin to protect against titania-induced neurotoxicity and the role of the Keap1/Nrf2/ARE signaling pathway. One group of animals were treated with Titania (0.045 and 0.075 g/rat) alone while the other with added melatonin (1 mg/kg and 3 mg/kg) and behavioral alterations were assessed using OFT (open field test). Neurochemical and histopathological changes were also studied in the hippocampus by analyzing kelch ECH associating protein 1 (Keap1), nuclear factor erythroid 2-related factor 2 (Nrf2), and antioxidant response element (ARE). It was seen that the animals with added Melatonin had improved behavioral scores in the OFT, like anxiety and motor dysfunction triggered by TiO2. Melatonin also reduced lipid peroxidation, ROS, GSSG, IL1ß, TNFα, Bax, and Keap1 levels, but boosted GSH, GPx, GR, SOD,IL10,IL4, Bcl2, Nrf2, and ARE levels and improved quadruple mitochondrial enzyme complex activity in titania-treated animals. Histopathological examination showed melatonin induced cytoprotection against vacuolization and necrosis in granular cells of DG and pyramidal cells of CA1 area of the hippocampus. In our study, pretreatment with melatonin reduced titania-induced neurotoxicity in the hippocampus through a mechanism potentially mediated by the Keap-1/Nrf2/ARE pathway.

Improved Healing of Colonic Anastomosis with Allotransplantation of Axillary Skin Fibroblasts in Rats.

Objective: Colonic anastomosis is associated with serious complications leading to significant morbidity and mortality. Fibroblasts have recently been introduced as a practical alternative to stem cells because of their differentiation capacity, anti-inflammatory, and regenerative properties. The aim of this study was to evaluate the effects of intramural injection of fibroblasts on the healing of colonic anastomosis in rats. Materials and Methods: Inbred mature male Wistar rats were used in this experimental study (n=36). Fibroblasts were isolated from the axillary skin of a donor rat. In the sham group, manipulation on descending colon was done during laparotomy. A 5 mm segment of the colon was resected, and end-to-end anastomosis was performed. In the control group, 0.5 ml of phosphate buffer saline (PBS) was injected into the colonic wall and in the treatment group, 1×106 fibroblasts were transplanted. Following euthanasia on day 7, intra-abdominal adhesion, leakage and peritonitis were evaluated by necropsy. Mechanical properties were assessed using bursting pressure and tensile tests. Inflammation, angiogenesis, and collagen deposition were examined histopathologically. Results: The mean scores for adhesion and leakage were decreased in the treatment group versus control samples. Lower infiltration of inflammatory cells was observed in the treatment group (P=0.03). Angiogenesis and collagen deposition scores were significantly increased in the fibroblast transplanted group (P=0.03). Tensile mechanical properties of the colon were significantly increased in the treatment group compared to the control sample (P=0.01). There was no significant difference between the control and treatment groups in terms of bursting pressure (P=0.10). Positive weight changes were found in sham and treatment groups, but the control rats lost weight after 7 days. Conclusion: The results suggested that allotransplantation of dermal fibroblasts could improve the necroscopic, histopathological, and biomechanical indices of colonic anastomosis repair in rats.

Allotransplantation of ascorbic acid-treated fibroblasts improves healing of excisional cutaneous wound in diabetic rats.

The purpose of this study was to evaluate the effects of ascorbic acid (AA)-treated fibroblasts transplantation on excisional diabetic wound healing. An excisional wound was created between the shoulders of streptozotocin-induced diabetic rats. On day three, 1 ml of PBS, 1 × 106 intact homologous fibroblasts, and 1 × 106 fibroblasts treated with 50 µM AA were injected subcutaneously around the wound edges in control, treatment-1 and treatment-2 groups, respectively. In the sham group, the wound was left intact. Wound area was measured by planimetry. On day 15, samples were harvested for histopathological examination and hydroxyproline content. Wound area in treatment-1 and - 2 groups was significantly decreased compared to other groups, on days 11 and 15. The hydroxyproline content was significantly lower in the control group compared to the other groups. Histopathology revealed significant increases in the number of neovessels, macrophages, lymphocytes and fibroblasts in the treatment-2 group compared to the other groups. Trichrome staining showed the highest level of collagen deposition and orientation in the treatment-2 group. In conclusion, allotransplantation of 50 µM AA-treated fibroblasts could result in progressive healing and improved reparative indices of excisional dermal wound in diabetic rats.

Evaluation of cardiovascular biomarkers and histopathological alterations in cattle naturally infected by Babesia bigemina.

Hemolytic anemia and secondary hypoxia are characteristics of naturally occurring Babesia bigemina infection in cattle. The anemic phase comes with cardiovascular insufficiency due to hypoxia-induced system dysfunction; but to date there is no description of cardiac damage in the infected animals. Therefore, this study was undertaken to investigate hematological parameters, biomarkers of cardiac function and D-dimer in 13 cattle infected with B. bigemina which were unresponsive to standard treatment. The animals were necropsied and the cardiac tissue was examined for histopathologic alterations. A significant parasitemia burden-dependent increase in the cardiac biomarkers and D-dimer level were recorded in the infected cattle compared to the control animals. Thrombocytes count was also significantly lower in the infected animals than the control. Both macroscopic and microscopic hemorrhage, mononuclear infiltrates, and myocardial necrosis were the evident histopathologic findings. These findings suggest that B. bigemina infection can potentially induce cardiac dysfunction in cattle. Furthermore, mechanistic studies should be conducted to understand the mechanisms beyond cardiac complications.

Evaluation of bilateral vasocystostomy for canine sterilization.

This study was aimed to evaluate canine vasocystotomy as a testosterone-preserving method of sterilization and investigate its potential post-operative complications. Five healthy adult male dogs underwent surgical procedure to transplant vasa deferentia to the urinary bladder. Under general anesthesia, caudal abdomen was opened and both vasa deferentia were ligated and transected. Then, the proximal free ends were sutured to mucosal layer of urinary bladder on its cranio-dorsal aspect. Serum testosterone level was measured on a weekly basis. Six-week postoperative assessments were performed including semen and urine sampling, ultrasound, contrast vasography, and tissue sampling. Statistical analyses revealed no significant differences in serum testosterone levels compared to its baseline value. Along with non-motile and broken spermatozoa, no cast or crystals were observed in urine samples. Semen analyses revealed azoospermia. No vasal obstruction or contrast leakage was observed in vasographs indicating bilateral patency in all dogs. Normal thickness of the bladder was found in ultrasounds. Histopathology showed normal testicular architecture and no inflammatory response was found in bladder or vas deferens following vasal transplantation. No significant change was found in testicular volume at the end of the study. This study suggested that vasocystostomy could be considered as an alternative method for canine sterilization with no significant changes in the testosterone concentrations and no evidence of postoperative complications. The preservation of testosterone could be regarded as an advantage and makes this approach favorable compared to the routine methods of sterilization especially for herding and guard dogs, because it prevents overpopulation while maintains the functionality.

Histopathological assessment of protective effects of selenium nanoparticles on rat hepatocytes exposed to Gamma radiation.

Gamma radiation are used in many medical and technical applications, however, it is one of the most dangerous kinds of radiation and can be harmful to the body. The present study was designed to clarify the protective effects of the selenium supplementation as selenium nanoparticle and selenite selenium in rat liver against Gamma irradiation with different intensities of 2.00 and 8.00 Gy. A total number of 45 healthy male Wistar rats were randomly divided into nine groups of five each. The radiation procedure was carried out in the Cobalt 60 equipment in Omid hospital, Urmia. The animals were simultaneously immobilized in a transparent acrylic plate and exposed to different intensities of 2.00 and 8.00 Gy radiations on day 7th and 14th of the experiment. After 72 hr after the last radiation, the animals were euthanized, and blood and liver tissue were collected. Histological analyses revealed the radiation-induced hepatic injury in rats, which included vacuolated cytoplasm, liver necrosis, fibrosis, and vascular lesions followed by a significant increase in alanine transaminase, alanine transaminase, alkaline phosphatase, and Gamma-glutamyl transferase. Selenium nanoparticles bear a more potent antioxidant effect in comparison with selenium selenite and can effectively protect the liver cell against Gamma radiation at a dose of 8.00 Gy.

Black proventriculus in Cobb 500 broiler breeders.

The black proventriculus is a necropsy finding sign observed during post-mortem examinations of dead young chicks up to 10 days of age and due to variation in color of the affected proventriculus organ, it may be ignored in some cases, in particular when the disease occurs simultaneously with other well-recognized infections of two weeks old chicks. In late January 2018, several live and dead chicks with a history of an unusual increase in mortality showing no clinical signs but sudden dead were referred to the poultry clinic of Faculty of Veterinary Medicine, Urmia University, Urmia, Iran. General routine postmortem examination revealed color changes (light green to dark green color) in the wall of proventriculus together with some degrees of congestion in the lungs and liver of the dead chicks. Microbiology tests revealed that the causative agent was Escherichia coli strain O142 and histopathological examination indicated that the color changing of the organ was due to necrosis of glandular epithelium with locally extensive or diffuse hypercellularity by lymphocytes and macrophages with hemorrhages. In conclusion, black proventriculus due to E. coli strain O142 was responsible for the sudden death of young chicks and based on antibiogram, selective antibiotics should be used for the treatment of the affected flocks.

The effects of crocin, mesalazine and their combination in the acetic acid-induced colitis in rats.

Crocin, as a carotenoid compound of saffron, exerts a potent antioxidant property. Mesalazine is frequently used in the treatment of ulcerative colitis. This study investigated the effects of separated and combination treatments with crocin and mesalazine in a rat model of ulcerative colitis. Ulcerative colitis was induced by intra-colonic administration of acetic acid (4.00%, 1.00 mL) at 8 cm proximal of the anus. Normal saline, acetic acid, crocin (5.00, 10.00 and 20.00 mg kg-1), mesalazine (100 and 300 mg kg-1) and crocin (5.00 mg kg-1) plus mesalazine (100 mg kg-1) were administered after induction of colitis for eight days. Body weight, organosomatic index (OSI), macroscopic and microscopic evaluations of colon and measurement of malondialdehyde (MDA), superoxide dismutase (SOD), tumor necrosis factor-alpha (TNF-α) contents of colon tissue were determined on day eight after induction of colitis. Crocin (10.00 and 20.00 mg kg-1), mesalazine (300 mg kg-1) and crocin (5.00 mg kg-1) plus mesalazine (100 mg kg-1) significantly (p < 0.05) improved body weight and OSI and reduced macroscopic and microscopic scores. These treatments also significantly (p <0.05) recovered the increased levels of MDA and TNF-α as well as the decreased level of SOD in colon tissue. Crocin and mesalazine did not produce significant effects in intact rats. Based on the results, it is concluded that crocin and mesalazine produced protective effects on colon tissue via antioxidant and anti-inflammatory actions. In addition, a synergistic effect was observed between crocin and mesalazine in attenuating ulcerative colitis.

Hemp seed/evening primrose oil affects expression of STAT3, IL-17, and FOXP3+ in experimental autoimmune encephalomyelitis.

T helper (Th)-17 mediate inflammation in both peripheral tissues and the central nervous system. Signal transducer and activator of transcription factor3 (STAT3) is required for Th-cell pathogenicity and its activation in the brain has been demonstrated during the acute phase of experimental autoimmune encephalomyelitis (EAE) through the mammalian target of rapamycin (mTOR) signaling. Rapamycin (RAPA), an inhibitor of mTOR, can drive Forkhead box P3 (FOXP3+) induction as a regulatory factor. The aim of this study was to determine the effects of hemp seed/evening primrose oils (HSO/EPO) supplement on the expression of FOXP3+, STAT3, and interleukin (IL)-17 genes in EAE lymph nodes. EAE was induced by myelin oligodendrocyte glycoprotein peptide in mice, and then the mice were assigned to three treatment groups compared to two control groups (EAE and naive). The histological findings of the spinal cord were evaluated. To determine the expression of FOXP3+, STAT3, and IL-17 genes in the lymphocytes, qRT-PCR was used. Our results showed that EAE severity was reduced in HSO/EPO mice by reducing the expression of STAT3 and IL-17 genes and increasing the expression of FOXP3+ gene, which was confirmed by slight inflammation in the spinal cord. Histological findings showed a significant improvement in the HSO/EPO group. Our findings suggest that the HSO/EPO treatment can be used to ameliorate the demyelination of spinal cord, which was confirmed by immunological and histological findings.

Upregulation of FSHR and PCNA by administration of coenzyme Q10 on cyclophosphamide-induced premature ovarian failure in a mouse model.

Cyclophosphamide (CTX) has been broadly used in the clinic for the treatment of autoimmune disorders and ovarian cancer. The process of chemotherapy has significant toxicity in the reproductive system as it has detrimental effects on folliculogenesis, which leads to an irreversible premature ovarian failure (POF). Coenzyme Q10 (CoQ10) has positive impacts on the reproductive system due to its antioxidant properties, protecting the cells from free-radical oxidative damage and apoptosis. However, little is known about the possible synergistic effect of CTX and CoQ10 on the expression of genes involved in folliculogenesis, such as proliferation cell nuclear antigen (PCNA) and follicle-stimulating hormone receptor (FSHR). A total of 32 NMRI mice were applied and divided into four groups, including healthy control, CTX, CTX + CoQ10, and CoQ10 groups. The effects of CoQ10 on CTX-induced ovarian injury and folliculogenesis were examined by histopathological and real-time quantitative reverse transcription-polymerase chain reaction analyses. The rates of fertilization (in vitro fertilization), embryo development, as well as the level of reactive oxygen species (ROS) in metaphase II (MII) mouse oocytes after PMSG/HCC treatment were also assessed. Results showed that the treatment with CTX decreased the mRNA expression of PCNA and FSHR, IVF rate, and embryo development whereas the application of CoQ10 successfully reversed those factors. CoQ10 administration significantly enhanced histological morphology and decreased ROS levels and the number of atretic follicles in the ovary of CTX-treated mice. In conclusion, it seems that the protective effect of CoQ10 is exerted via the antioxidant and proliferative properties of this substance on CTX-induced ovarian damage.

Comparison Therapeutic Effects of Ciprofloxacin, Silver Nanoparticles and Their Combination in the Treatment of Pseudomonas keratitis in Rabbit: An Experimental Study.

Pseudomonas aeruginosa is one of the most common causes of keratitis. The current study was done to evaluate the therapeutic effects of antibacterial combinations with Silver nanoparticles (Ag-NPs) and Ciprofloxacin in experimental Pseudomonas keratitis. Sixty four New Zealand rabbits were prepared. All rabbits were randomly categorized into eight groups (each group containing eight rabbits): Control +, Control -, Ciprofloxacin, Ag-NPs, Ciprofloxacin plus Betamethasone, Ag-NPs plus Betamethasone, Ciprofloxacin plus Ag-NPs, and Ciprofloxacin plus Ag-NPs plus Betamethasone. Twelve hours after bacterial inoculation into the cornea, the eyes were examined daily to evaluate the number of days of ocular discharge and blepharospasm. Also, after 108 and 204 h, first grading of corneal opacity was done and then four rabbits of each groups were euthanized for bacterial count. The results showed that the means of days of blepharospasm, ocular discharge, and bacterial counts (log CFU mL-1) were significantly different in the treatment groups at 108 and 204 h (P <0.0005, ANOVA). According to Tukey's test, Ciprofloxacin plus Ag-NPs plus Betamethasone group was significantly less than Control +, Ag-NPs, and Ag-NPs plus Betamethasone groups for these variables (P < 0.05). The mean rank of opacity scores was significantly different between treatment groups (P = 0.01, Kruskal-Wallis). Mann-Whitney U-test revealed that Ciprofloxacin plus Ag-NPs plus Betamethasone group had significantly better score than Control +, Ag-NPs, and Ag-NPs plus Betamethasone groups (P < 0.05). It seems Ag-NPs can be an appropriate adjuvant for Ciprofloxacin, but due to the results they can't be an alternative for Ciprofloxacin to treat Pseudomonas keratitis.

Regulatory effects of hemp seed/evening primrose oil supplement in comparison with rapamycin on the expression of the mammalian target of rapamycin-complex 2 and interleukin-10 genes in experimental autoimmune encephalomyelitis.

The mammalian target of rapamycin (mTOR) signaling plays a critical role in lipid synthesis and immune responses. The T regulatory cells (Treg) as suppressor of T cells, are a subset of T cells that modulate the immune system, maintain tolerance, and prevent autoimmune diseases.. The interleukin (IL) -10 derived from the Treg and T helper (Th) 2 is an anti-inflammatory cytokine in multiple sclerosis (MS) and experimental autoimmune encephalomyelitis (EAE). Due to the exclusive roles of rapamycin (RAPA) in mTOR inhibition, we evaluated the regulatory effect of the hemp seed oil/evening primrose oil (HSO/EPO) supplement in comparison with RAPA in EAE. EAE was induced by using myelin oligodendrocyte glycoprotein peptide and complete freund's adjuvant (CFA) in C57BL/6 mice, total mRNA was extracted from local lymph nodes and real-time polymerase chain reaction was used to evaluate the expression level of the rapamycin-insensitive companion of mTOR complex 2 (RICTOR) and IL-10 genes. The expression of IL-10 and RICTOR genes were significantly increased in HSO/EPO group. In contrast with RAPA groups, histological findings have shown that the HSO/EPO treated group remarkably reduced cell infiltration and promoted remyelination. The EPO/HSO has beneficial effects on the repair of myelin, which was confirmed by immunological and histological findings.

In vivo toxicity of orally administrated silicon dioxide nanoparticles in healthy adult mice.

The increasing use of silica nanoparticles (SiNPs) in various applications including industrial, agriculture, and medicine has raised concerns about their potential risks to human health. Various nanotoxicity researches have been done on the assessment of SiNPs' toxic effects; however, a few in vivo investigations exist. In this investigation, an in vivo study was done in order to evaluate the oral toxicity of SiNPs. The biochemical levels of 19 different serum parameters were assessed. Moreover, the histopathological changes have been examined as well. We showed that SiNPs with diameters of 10-15 nm in size can cause significant changes in albumin, cholesterol, triglyceride, total protein, urea, HDL, and LDL as well as in alkaline phosphatase and aspartate aminotransferase activity. In addition, histopathological examinations demonstrated that SiNPs have toxic effects on various tissues including liver, kidney, lung, and testis.

Histopathological changes caused by the nymph stage of Linguatula serrata in the mesenteric lymph nodes of goats.

Linguatula serrata (Fröhlich, 1789) is a ubiquitous parasite species belonging to pentastomids, which has been reported frequently from domestic herbivores of Iran. This study was carried out to elucidate the pathological changes caused by the nymphal stage of L. serrata in the mesenteric lymph nodes (MLNs) of goats. Jejunal and ileal MLNs of slaughtered goats infected with L. serrata were collected and fixed in 10% neutral formal saline. Histopathological changes included follicular necrosis accompanied by a loss of lymphocytes and oedematous cystlike spaces. Granulomatous lesions were formed by mononuclear cell infiltration and necrotic tissue. Some lymphocytes were apoptotic with chromatolysis. A more frequent vascular abnormality was vasculitis. These findings show that larval migration is the main factor to initiate trauma and cause irritation in the MLNs of goats. Since goats are habitual intermediate hosts in Iran, rigorous inspection of the MLNs of slaughtered goats is recommended in order to decrease the chance of human infection through the consumption of raw visceral organs of goats.

Toxicity of ZnO nanoparticles in healthy adult mice.

In this study, we extended previous work to evaluate the oral toxicity of ZnO nanoparticles and their possible effects on different serum-elements and sexual hormones in the mouse. The histopathological changes have also been examined. Significant recorded increases in alanine aminotransferase and aspartate aminotransferase activity in all mice exposed to ZnO nanoparticles suggest that these nanoparticles can cause hepatic injury. Hepatocyte necrosis and other pathological observations also confirmed liver damage. Moreover, Glomeruli segmentation, hydropic degeneration in epithelial cells, necrosis of epithelial cells in tubules and swelling in epithelial cells of proximal tubules were found in all kidney tissues, which demonstrated that ZnO nanoparticles have severe toxicological effects on kidney. Serous inflammation, severe hyperemia in alveoli, and edema were observed as pathological findings in the lung which suggest that the lung is the third target tissue of the ZnO nanoparticles.

The occurrence of helminth parasites in the gastrointestinal of catfish (Silurus glanis Linnaeus 1758) from the Zarrine-roud river, Iran.

This investigation was undertaken to verify the prevalence of helminths parasites in the gastrointestinal tract of catfish. A total number of 116 catfish (Silurus glanis) were collected from Zarrine-roud river and examined for helminths. Fish were examined after washing contents of gastrointestinal tract and observed for the presence of helminths using a stereo microscope and a light microscope. Results indicated that 18.96% of the examined catfish were infected with digenean trematodes including Orientocreadium siluri (27%), Crowcrocoecum skrjabini (39%), and cestode Bothriocephalus gowkongensis (34%). All the parasites were found in the intestine. Mid-gut followed by foregut appeared to be the most commonly infected parts of the alimentary tract of hosts. The results showed that there was a significant correlation between infection rate, catfish body size, and weight (P < 0.05).