In vitro Antibacterial Activity and Wound Healing Effects of Achillea millefolium Essential Oil in Rat.

Objectives: In this study we aimed to evaluate the in vitro antibacterial activity and wound healing properties of Achillea millefolium essential oil (AMEO) in full-thickness wound model in rat. The antibacterial activity of AMEO was evaluated against Staphylococcus aureus and Pseudomonas aeruginosa using the broth dilution method. Methods: The 2 cm × 2 cm full-thickness excisional wounds were created on the back of animals. Topical therapy was applied twice a day using 1%, 2%, and 3% w/w AMEO ointments, and the measurement of the wounds area was carried out every 3 days, after that the wound closure percentage was calculated in these days. Hydroxyproline content and histopathological evaluation of wound tissue samples were carried out on day 7 and 14 post wounding. Eucerin was used for the treatment of vehicle control group and negative control group received no treatment. Results: Our results revealed the bacteriostatic activity of AMEO against S. aureus and P. aeruginosa. Wound healing activity evaluation of AMEO showed the significant increase (p < 0.05) in the wound closure percentages in rats treated with AMEO 1% and 2% comparing to those of non-treatment group. In addition, hydroxyproline contents of tissue significantly (p < 0.01) increased in AMEO 1% and 2% comparing to non-treatment group. Histopathological evaluations of wound tissue samples on day 7 and 14 demonstrated higher accumulation of collagen fibers, reduction of edema and inflammation and also formation of tissue appendages in 1% and 2% AMEO treated groups in comparison with non-treatment group. Conclusion: The results of this study indicated that AMEO has the potential to be used as a safe and effective wound healing agent.

Vitamins A, C, and E Exert Anti-apoptotic Function in the Testis of Rats After Exposure to Zinc Oxide Nanoparticles.

Some reports emphasize that zinc oxide nanoparticles (ZnO NPs) are detrimental to the reproductive organs of animals. As such, this research aimed at exploring the apoptotic potential of ZnO NPs on testis along with the beneficial role of Vitamins (V) A, C, and E against ZnO NP-induced damage. To this aim, a population of 54 healthy, male Wistar rats were used in this work and then assigned into nine groups of 6 rats as G1: Control 1 (Water); G2: Control 2 (Olive oil); G3: VA (1000 IU/kg), G4: VC (200 mg/kg), G5: VE (100 IU/kg), G6: ZnO NPs exposed animals (200 mg/kg); and G7, 8 and 9: ZnO NPs-exposed animals that were pre-treated with either VA, C, or E. Apoptosis rates were estimated by measuring the level of apoptotic regulatory markers including Bcl-2-associated X (Bax) and B-cell lymphoma protein 2 (Bcl-2) using western blotting and qRT-PCR assays. The data indicated that ZnO NPs exposure elevates the level of Bax protein and gene expression, whereas the protein and gene expression of Bcl-2 was reduced. Further, the activation of caspase-3,7 occurred after exposure to ZnO NPs, while the above alterations were significantly alleviated in the rats that were co-treated with VA, C, or E and ZnO NPs relative to the rats in the ZnO NPs group. In summary, VA, C, and E exerted anti-apoptotic functions in the testis of rats following administration of ZnO NPs.

Neospora caninum and Toxoplasma gondii infections in one-humped camels (Camelus dromedarius) in central desert of Iran.

The protozoan parasite Neospora caninum infects carnivores as definitive and a wide range of mammals as intermediate hosts. This parasite is regarded as an important cause of abortion in cattle worldwide, causing significant economic losses. Although there is serological evidence of infection in Old World camelids, the significance of N. caninum in these animal species is still poorly understood. The aim of this study was to use molecular and histological methods to detect N. caninum in the blood and tissues of 100 slaughtered one-humped camels (Camelus dromedarius) in Iran. For this, genomic DNA was extracted from blood, brain, portal lymph node and liver of the camels, and nested-PCR assay followed by sequencing were performed. Besides, paraffin-embedded tissue sections were stained with hematoxylin and eosin (H&E) and studied microscopically. In addition, immunohistochemical staining for N. caninum was attempted on brain samples with positive PCR results. All animals were tested for antibodies against N. caninum and Toxoplasma gondii by whole tachyzoite-agglutination tests. N. caninum DNA was detected in blood, brain, and portal lymph node, but not in the liver of two (2%) camels. Histopathological examination revealed cysts resembling N. caninum in brain samples of one of these camels; however, immunohistochemical staining for N. caninum and T. gondii did not allow a morphological identification. IgG antibodies to N. caninum and T. gondii were detected in 36% and 35% of the camels, respectively. This study provides the first insight into direct detection of N. caninum in C. dromedarius in Iran. Further molecular studies on aborted fetuses, stillborn animals and cases of perinatal mortality are needed to understand the possible involvement of N. caninum in cases of reproductive failure. As the definitive hosts of N. caninum are domestic and wild canids, producers should be advised to monitor and limit exposure of their camelids to these species and their feces.

Protective efficacy of dark chocolate in letrozole-induced ovary toxicity model rats: hormonal, biochemical, and histopathological investigation.

OBJECTIVE: To assess the protective effect of dark chocolate (DC) on the letrozole-induced rat model of polycystic ovary syndrome (PCOS). METHODS: In this experimental study, 32 female Wistar rats, weighing (200 ± 20) g, were randomly categorized into 4 groups including control, letrozole (1 mg·kg·d), metformin (500 mg·kg·d) along with letrozole, and DC (500 mg·kg·d) along with letrozole for 28 d by oral gavage. Twenty-four hours after the last supplementation, direct blood sampling was taken from the heart to obtain blood serum for evaluation of sex hormones and gonadotropins, oxidative parameters, inflammatory cytokines, and ovarian tissue was examined for histology. RESULTS: The DC treatment significantly improved PCOS signs, as demonstrated by the significant restoration of ovarian morphology and physiological functions as compared with the letrozole group. DC treatment also decreased ovarian interleukin-1ß and tumor necrosis factor-α levels and improved total oxidative/antioxidative status as compared with the letrozole group. CONCLUSIONS: Treating the animals with DC could alleviate the PCOS symptoms and reduced the toxic effects of letrozole in the ovary. This effect may mediate through antioxidant and antiinflammatory properties.

Gelatin-starch composite coating containing cucumber peel extract and cumin essential oil: Shelf life improvement of a cheese model.

In this study, the effects of gelatin-starch (GS) composite coating containing cucumber peel extract (CPE) and cumin essential oil (CEO) were evaluated on the shelf life enhancement of ultrafiltered (UF) cheese during 56 days of storage under refrigerated conditions. The obtained hydroethanolic CPE by the microwave method showed the best results in terms of the total phenolic content, reducing power, 2,2'-diphenyl-1-picrylhydrazyl (DPPH) activity, and 2,2-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) radical scavenging activity compared to the immersion and ultrasound methods. The studied treatments were as follows: Control (C), GS, CPE, CEO, GS-CPE, GS-CEO, and GS-CPE-CEO. Scanning electron microscopic surface morphology of treated cheese samples showed the formation of a firm, integrated, flawless, and homogenous layer on the cheese slices of the GS-CPE-CEO treatment. All treatments significantly (p ≤ .05) decreased the total viable count, psychotropic bacteria, and yeast-mold population compared to the control group. Adding CEO and/or CPE to GS significantly (p ≤ .05) controlled undesirable changes in physical characteristics, such as weight, color, and hardness of the cheese slices. Throughout storage time, the coated cheese slices showed more stable chemical features in comparison to the uncoated cheese samples in terms of moisture, lipid oxidation, pH, and titratable acidity (TA). Sensory evaluation of the preparations showed that the GS coating containing CPE and CEO significantly (p ≤ .05) had pleasant effects on the sensory features (taste, odor, texture, and overall acceptability) of the cheese samples during storage time. It was concluded that composite coating of GS containing CPE and CEO could improve the microbial, physical, chemical, and sensory features of ultrafiltration (UF) cheese during refrigerated storage.

The Restoring Effect of Human Umbilical Cord-Derived Mesenchymal Cell-Conditioned Medium (hMSC-CM) against Carbon Tetrachloride-Induced Pulmonary Fibrosis in Male Wistar Rats.

Objective: Pulmonary toxicity induced by CCl4, a model of idiopathic pulmonary fibrosis (IPF), leads to tissue remodeling and inflammation. Human umbilical cord mesenchymal cell-conditioned medium (hMSC-CM) is a potent anti-inflammatory, antioxidative, and antifibrotic agent. Methods: Forty male Wistar rats were assigned to the control (C), olive oil control (C.O) (hMSC-CM), control (C.Ms), fibrosis (fb), and fibrosis with hMSC-CM (f.Ms) treatment groups. The groups C, C.O, and C.Ms received PBS (200 µl), olive oil (1 ml/kg), and hMSC-CM (100 µg protein/kg), respectively. The fibrosis group was administered with only CCl4 (1 ml/kg). The last group, f.Ms was treated with CCl4 (1 ml/kg) and 100 µg protein/kg IV hMSC-CM. While the treatment with olive oil and CCl4 was performed for 2 days/week from the first week for 12 weeks, the treatment with PBS and hMSC-CM was carried out 2 days/week from week 4th to week 12th. The effect of the UC-MSC culture medium treatment on the lung was evaluated by assessing lysyl oxidase (LOX), tumor necrosis factor-alpha (TNF-α), and transforming growth factor-ß1 (TGF-ß1) genes, and proteins expression by real-time RCR and western blotting, respectively. Results: Lysyl oxidase (LOX), tumor necrosis factor-alpha (TNF-α), transforming growth factor-b1 (TGF-ß1), malondialdehyde (MDA), and oxidative stress levels were markedly higher in the fibrosis group than in the control groups (p ≤ 0.001). Additionally, glutathione (GSH) in the fibrosis group was markedly lower than those in the control groups (p ≤ 0.001). Fibrosis in the UC-MSC treatment group had milder histopathological injuries than in the fibrosis group. Conclusion: hMSC-MSC as a strong anti-inflammatory, antioxidative, and antifibrotic decreases the level of oxidative stress, proinflammatory cytokines, and MDA causing a restoring effect against CCl4-induced pulmonary fibrosis.

Treatment with human umbilical cord blood serum in a gentamicin-induced nephrotoxicity model in rats.

Sold under the brand name of Garamycin, gentamicin (GM) is an antibiotic in the category of aminoglycoside, that although does have many antibacterial properties, owing to several side effects, its consumption is confined. The current study is aimed at gauging the protective influences of human umbilical cord blood serum (hUCBS) on nephrotoxicity which is induced by GM. In this regard, in the present experimental design, twenty-eight male Wistar rats with the weights of 220 ± 20 g were categorized randomly into 4 groups of seven. The groups included GM (100 mg/kg), control as well as hUCBS at doses of one and two percent together with GM (100 mg/kg) for ten days in an intraperitoneal manner. Blood sampling was collected from the heart directly 24 h after the final injection for obtaining blood serum; the parameters of C-reactive protein (CRP), total oxidant status (TOS), interleukin (IL)-6, lactate dehydrogenase (LDH), total antioxidant capacity (TAC), creatinine (Cr), blood urea nitrogen (BUN), blood serum glutathione (GSH) were gauged in blood serum samples to evaluate renal function. Moreover, for histology, an examination of kidney tissue was performed. In comparison to those of the GM group, in the treatment group, hUCBS significantly decreased the levels of BUN, Cr, LDH, TOS, IL-6, and the CRP levels, and significantly increased the TAC and GSH levels. It was revealed that the treatment of the animals with hUCBS culminates in the reduction of GM' toxic impacts on the kidney.

Cuminum cyminum L. Essential Oil: A Promising Antibacterial and Antivirulence Agent Against Multidrug-Resistant Staphylococcus aureus.

Cuminum cyminum L. (cumin) is valued for its aromatic and medicinal properties. There are several reports of antibacterial activity of C. cyminum essential oil (CcEO). Accordingly, the present study was conducted to investigate the mechanism(s) of action of the CcEO against multidrug-resistant (MDR) Staphylococcus aureus. Therefore, 10 S. aureus MDR isolates, obtained from different sources, were selected based on the antibiotic susceptibility patterns and the Clinical and Laboratory Standards Institute definition and subjected to the examinations. Our results exhibited promising bacteriostatic and bactericidal properties of the CcEO. The minimum inhibitory concentration (MIC) and the minimum bactericidal concentration values ranged from 5 to 10 and 10 to 20 µL ⋅ mL-1, respectively. Scanning electron microscope was used to assess the bacterial cell structure and morphology after the induction with 1/2 MIC concentration of the CcEO. The observed morphological changes appeared to be deformation of the cell membrane and destruction of the cells. In the case of quorum sensing inhibitory potential, treatment of S. aureus isolates with the sub-MIC concentrations (1/2 MIC) of the CcEO significantly reduced the hld expression (3.13-fold downregulation), which considerably controls S. aureus quorum-sensing accessory regulator system. Another virulence factor influenced by the CcEO was the polysaccharide intercellular adhesion production system, as an important component of cell-cell adhesion and biofilm formation. Consequently, the expression level of the intercellular adhesion (ica) locus in the S. aureus cells was examined following treatment with CcEO. The results showed significant decrease (-3.3-fold) in ica expression, indicating that the CcEO could potentially interfere with the process of biofilm formation. Using the ethidium bromide efflux inhibition assay, the S. aureus NorA efflux pump was phenotypically but not genotypically (in quantitative polymerase chain reaction assay) affected by the CcEO treatment. Using gas chromatography-mass spectrometry analysis, cuminic aldehyde (38.26%), α,ß-dihydroxyethylbenzene (29.16%), 2-caren-10-al (11.20%), and γ-terpinene (6.49%) were the most detected compounds. The antibacterial and antivirulence action of the CcEO at sub-MIC concentrations means that no microbial resistance will be promoted and developed after the treatment with this agent. These findings revealed that the CcEO is a promising antibacterial agent to control infections caused by the MDR S. aureus strains.

Hepatic Capillaria hepatica (Bancroft, 1893) infection in cat (Felis catus)-histopathological findings and first report from Iran.

Capillaria hepatica (syn. Calodium hepaticum) is a globally distributed nematode with a high affinity to the liver of a wide range of mammalian hosts, including humans. Documented reports of the nematode in cats and associated histopathology are rare. Here, we describe a case of C. hepatica infection in a 5-year-old male stray cat from Iran. At post-car accident necropsy, all body parts appeared normal except for the liver, in which a few yellowish-white granulomatous nodules were observed through the capsule and in the organ. Histopathological examination of the tissue revealed a large number of clustered parasite eggs in the parenchyma. The barrel-shaped, un-embryonated eggs (55.19 × 28.37 µm), with inconspicuous caps at both ends, were covered with striated shells. The presence of ova in the liver tissue had resulted in the development of hepatic inflammation with hepatocellular necrosis associated with the development of multifocal granulomas. As predators of small rodents, the cats might have a significant role in the epidemiology of C. hepatica. Infection of hosts through ingestion of embryonated eggs in contaminated water, food, or soil is of major importance in the epidemiology of C. hepatica. Since the rare reports of feline infection have come mainly from accidental detection of the parasite, any hepatic disease presenting difficulties to find an etiological agent may virtually be associated with the infection with this little-known nematode.

Stability and antibacterial activity of Thymus daenensis L. essential oil nanoemulsion in mayonnaise.

BACKGROUND: There is a growing demand in the food industry for the replacement of synthetic preservatives with their natural alternatives. This has led to the development of novel methods such as encapsulation of plants essential oil with appropriate physicochemical stability, and antibacterial and organoleptic properties. This study aimed to prepare an optimal nanoemulsion of Thymus daenensis L. essential oil for use as a natural preservative in mayonnaise. RESULTS: The analysis of droplet diameter, polydispersity index, zeta potential, encapsulation rate, and intrinsic stability showed that out of nine T. daenensis essential oil-containing nanoemulsions, two preparations of A and B had high stability scores. In vitro antibacterial tests showed the adverse effect of Tween 80 volume on the antibacterial properties of nanoemulsions. One nanoemulsion (essential oil:Tween 80, ratio 1:1, 15 min sonication) was considered to be optimal based on its long-term stability and antibacterial effects on Salmonella Typhimurium, Escherichia coli, and Listeria monocytogenes. However, compared to the optimal nanoemulsion, the pure essential oil showed more antibacterial effects. The bacterial control in mayonnaise was close to equal for sodium benzoate (in maximum limit 1 g kg-1 ) and the optimal nanoemulsion (½ MIC) for 24 h. The optimal nanoemulsion achieved significantly higher sensory scores (taste, appearance, and mouthfeel) than the pure essential oil in mayonnaise (P < 0.05). CONCLUSION: The results demonstrated similar antibacterial effects for the optimal nanoemulsion and sodium benzoate. The optimal nanoemulsion, due to its desirable sensorial attributes, long-term stability, and slow release of volatile compounds, can be considered an appropriate alternative to synthetic preservatives.

Therapeutic effects of mesenchymal stem cells on cutaneous leishmaniasis lesions caused by Leishmania major.

OBJECTIVES: Leishmania major (L. major) is a cutaneous leishmaniasis causative agent. Current chemotherapeutic methods are not totally effective in treatment of this disease. The immunomodulation and tissue repairing capability of mesenchymal stem cells (MSCs), ease of isolation, detection and in vitro culture, have encouraged biologists to use MSCs for cell therapy in different infections such as cutaneous leishmaniasis. METHODS: BALB/c mice (6-8 weeks old) were infected with L. major then divided into four groups and treated with MSCs, Glucantime, Glucantime + MSCs, or PBS. Regression of lesions, potency of macrophages for phagocytosis, proliferation of immune cells against Leishmania soluble antigen, reduction of spleen parasite burden and healing of the lesions were evaluated on days 10, 20 and 30 of treatment. RESULTS: The results indicated that the mice intralesionally injected with MSCs showed significant regression in the lesions produced by L. major by day 30. Proliferation of splenocytes stimulated with SLA (soluble leishmania antigen) in vitro in MSC-treated mice on day 20 was significantly higher than in the other groups. The potency of phagocytosis in macrophages of mice treated with MSCs was significantly higher by day 30 and healing of the lesions in this group of mice showed more progress on histopathological examinations. Spleen parasite burden showed significant reduction in the mice treated with Glucantime + MSCs by day 30. CONCLUSIONS: The results showed that including MSCs in treatment of cutaneous leishmaniasis caused by L. major is a promising approach.

Bisphenol-A analogue (bisphenol-S) exposure alters female reproductive tract and apoptosis/oxidative gene expression in blastocyst-derived cells.

OBJECTIVES: One of the major endocrine-disrupting chemicals, bisphenol-S (BPS) has replaced bisphenol-A due to public health anxiety. The present study evaluated low dosage BPS effect on female reproductive potential, hormonal disruption, and gene expression pathways of blastocyst-derived cells. MATERIALS AND METHODS: NMRI female mice (5-6 weeks) in the estrous stage were chosen following vaginal smear examination for estrus cycle detection and BPS (0, 1, 5, 10, 50 and 100 µg/kg) was administrated subcutaneously for twenty-one consecutive days. After the last administration, blood, ovary tissue and oocytes were collected for further examination. RESULTS: BPS induced oxidative stress in ovarian tissue and reduced hormonal status, LH and FSH, even at low concentration. Furthermore, apoptosis was induced in blastocyst derived cells in BPS administrated mice groups even at low BPS concertation, however, P53 and E2f1 expression were downregulated in doses more than 50 µg/kg, which might indicate apoptosis pathway exchange from P53 dependent to p53 independent pathways. IVF outcome was negatively associated with blastocyst apoptosis gene expression, estrogen receptor beta (ERß) as well as oxidative status in ovaries. Finally, Stepwise regression indicated that E2f1, Nrf2, catalase (CAT), and malondialdehyde (MDA) could be chosen as predictor values for hatch percentage in IVF outcome. CONCLUSION: In summary, this study revealed BPS might have detrimental potential in the female reproductive system by oxidation induction and hormonal alteration as well as next generation blastocyst derived cells apoptosis induction. Further studies are recommended for public health assurance of BPS safety especially for female consumed products.

Effect of Rosa canina Distilled Water on Tamoxifen-treated Male Wistar Rats.

BACKGROUND AND OBJECTIVE: In spite of therapeutic effect of tamoxifen on the breast cancer, it has some side effects on the liver including non-alcoholic fatty liver disease. In this study the effects of Rosa canina distilled water on the tamoxifen-induced fatty liver and oxidative stress status in male rats were investigated. MATERIALS AND METHODS: Twenty four adult male Wistar rats were randomly divided into 4 groups of 6: 1st group: Untreated control rats (C), 2nd group (T): The rats received tamoxifen, 3rd group (T+R): Rats received tamoxifen and Rosa canina distilled water and 4th group (R): Rats received only Rosa canina distilled water. Tamoxifen at 1 mg kg-1/day was injected subcutaneously for 7 days and the rats received orally Rosa canina distilled water at 1 mL/rat/daily for 14 days. At the end of the study, animals were studied for serum biochemical parameters (glucose, lipid profile, BUN, creatinine, uric acid, urea, ALT, AST, ALP, total protein, bilirubin, oxidative stress indices, sperm analysis and histology of the liver. The data were analyzed with SPSS software version 20 and expressed as Mean±SD. RESULTS: Rosa canina distilled water improved liver enzyme and renal function indices which disturbed due to tamoxifen treatment. While tamoxifen enhanced lipid peroxidation, Rosa canina distilled water reduced it. In addition, tamoxifen reduced the mobility, morphology and viability of sperms, but the Rosa canina distilled water enhanced the sperm parameters. Histological results also confirmed the adverse effect of tamoxifen and the favorable impact of the Rosa canina distilled water on the liver structures of animals. CONCLUSION: Rosa canina distilled water could modulate tamoxifen-induced fatty liver as well as improving the sperm parameters.

Study of telomerase reverse transcriptase and uterine-ovarian-specific genes expression in the endometrial tissue of ovariectomized female Sprague-Dawley rats.

BACKGROUND: An in vivo study was carried out to study of telomerase reverse transcriptase and Uterine-Ovarian-specific genes expression in the endometrial tissue of ovariectomized female Sprague-Dawley rats. MATERIAL AND METHODS: Twenty-four female Sprague-Dawley rats divided into 4 groups of six rats. The first and second groups were ovariectomized and given tamoxifen and tamoxifen-loaded SLN respectively for six days continuously. Group 3 served as the untreated ovariectomized control group and group 4 was made up of untreated normal healthy rats. At the end of the study, the rats were sacrificed and study of the genes expression and serum zinc and copper were carried out. RESULTS: The results showed that the expression of TERT in the group treated with tamoxifen, and tamoxifen-loaded solid lipid nanoparticles, significantly decreased (p<0.001) compared with ovariectomized control group. The results also revealed that the treatment with tamoxifen-loaded solid lipid nanoparticles increased expression of UO-44 gene compared to ovariectomized control group, while there was no difference between tamoxifen treated and control group. CONCLUSIONS: Encapsulation of tamoxifen in solid lipid nanoparticles increased its targeting efficiency and improved the impact of the drug on the serum levels of some trace elements.