ABSTRACT
Antioxidants are commonly used for maturation, fertilization and early development of embryos. Melatonin as an antioxidant have been recently proven to be useful for the assisted reproductive technology. In the present study, we evaluated the roles of melatonin in the in vitro maturation, fertilization, development and also the gene expression of high mobility group box-1 (HMGB1) in the blastocysts. The immature oocytes of BDF1 mice were transferred to the media containing different doses of melatonin (10-6, 10-9, 10-12 M). The blastocysts that developed under in vitro fertilization from each group were stained to determine the cell number of embryos and analyzed to determine the expression level of HMGB1 by real-time PCR. The most effective doses of melatonin for maturation of oocytes were 10-6 and 10-12M (P<0.05). Fertilization rate, early development and the cell number of blastocysts were significantly higher in the group that treated with 10-12 M of melatonin comparing to the other groups. The HMGB1 expression decreased in groups that treated with 10-6M and 10-9M of melatonin and increased in the group that treated with 10-12 M of melatonin, but did not show a significant difference (pË0.05). From the results, it may be concluded that the melatonin could be effective when the embryos undergo maturation, fertilization and early developmental processes. The HMGB1 expression, as a marker of early development in mice embryos, increased in the groups that treated with low doses of melatonin
Subject(s)
Animals , Female , Mice , Blastocyst , Fertilization in Vitro , Embryonic Development , In Vitro Oocyte Maturation Techniques/instrumentation , Melatonin/adverse effects , Gene Expression , Cell Count/instrumentation , Reproductive Techniques, Assisted , Embryonic Structures , Antioxidants/administration & dosageABSTRACT
Background: Hypothyroidism is associated with dysfunction of the bone turnover with reduced osteoblastic bone formation and osteoclastic bone resorption. Mesenchymal stem cells [MSCs] secrete various factors and cytokines that may stimulate bone regeneration. The aim of this study was to determine the effects of MSCs-conditioned medium [CM] in hypothyroidism male rats after inducing bone defect
Methods: In this study, 24 male rats were randomly assigned to three groups: [I] hypothyroidism + bone defect [HYPO], [II] hypothyroidism + bone defect + CM [HYPO + CM], and [III] no hypothyroidism + bone defect [control]. Four weeks after surgery, the right tibia was removed, and immediately, biomechanical and histological examinations were performed
Results: The results showed a significant reduction in bending stiffness [32.64 +/- 3.99], maximum force [14.63 +/- 1.89], high stress load [7.59 +/- 2.31], and energy absorption [12.68 +/- 2.12] at the osteotomy site in hypothyroidism rats in comparison to the control and hypothyroidism + condition medium groups [p < 0.05]. There was also a significant decrease in the trabecular bone volume [3.86 +/- 3.88] and the number of osteocytes [5800 +/- 859.8] at the osteotomy site in hypothyroidism rats compared to the control and hypothyroidism + condition medium groups [p < 0.01 and p < 0.02, respectively]
Conclusion: The present study suggests that the use of the CM can improve the fracture regeneration and accelerates bone healing at the osteotomy site in hypothyroidism rats
Subject(s)
Animals, Laboratory , Culture Media, Conditioned , Hypothyroidism/veterinary , Osteotomy , Tibial Fractures , Fracture HealingABSTRACT
An adult native cock [Gallus gallus domesticus] referred to the aviary clinic with multiple different sizes of round dermal nodules. The bird died few days later, and was then submitted for further evaluation. Macroscopic and microscopic examinations as well as a PCR test were done to identify type and cause of the tumor. In histopathological assessment of biopsy specimen, it consisted of interlacing bundles of fibroblasts that orientated in different directions with plump or elongated spindle shaped nuclei and fairly abundant cytoplasm. At necropsy several large white nodules were implanted in lung and liver. Microscopically the proliferated fibroblastic cells were invaded to both organs, and were similar to those described for skin lesion. The tumor cells had immunoreaction for alpha smooth muscle actin, vimentin and S100 protein, whereas they were negative for desmin and pancytokeratin, suggesting a diagnosis of metastatic neuro-myofibroblastic sarcoma. A PCR test specific for avian leukosis virus subgroup J [ALV-J] confirmed the presence of that virus in tumor specimens. Sequencing and phylogenetic analysis showed a relatively low similarity in the LTR segment [90%] of the studied virus with other ALV-J strains. It might be the first report of cutaneous neuro-myofibroblastic sarcoma, potentiated to metastasis to other organs induced by ALV-J
ABSTRACT
BACKGROUND: In recent years, dietary fat supplementation has become a common practice and fat supplemented diets have had variable effects on animal performance and carcass characteristics.
OBJECTIVES: This study was conducted to determine the effect of increasing the levels of sesame oil on performance, rumen parameters and plasma lipid profile in finishing Chal lambs.
METHODS: Eighteen male lambs were fed with control [C; without sesame oil] and the diets contained low level [LSO; 2.5%] and high level [HSO; 5%] of sesame oil.
RESULTS: The addition of fat had no effect on dry matter intake [DMI], average daily gain [ADG], feed conversion ratio [FCR] and carcass weight. Treatments with sesame oil had no effect on rumen ammonia concentration and total protozoal count. Ruminal pH was numerically smaller [p=0.14] for control compared to lambs fed diets containing 2.5 and 5% sesame oil. Treatments had no effect on total VFA, acetate, butyrate, valerate and isovalerate concentrations. The propionate concentration decreased [p=0.04] by supplementation of sesame oil compared with the control. Total cholesterol [TC] and HDL increased and the tendency for VLDL and triglyceride [p=0.13] to increase was observed by treatment groups when compared with the control. Glucose concentration and LDL were not affected by treatment.
CONCLUSIONS: Sesame oil had no effect on the performance and carcass weight of lambs but there was decrease in blood cholesterol concentration.
ABSTRACT
Objective: To determine the presence of antibodies against phase II among slaughterhouse workers in Kerman, southeast of Iran.Methods:sorbent assay using phase II Coxiella burnetii as the antigen [kit (Virion\Serion, Wurzburg, Germany) according to the manufacturer’s protocol].Results:The antibody titers of the serum samples were measured by enzyme-linked immuno Conclusions: Our findings suggest that slaughterhouse workers in Kerman area have a higher risk of infection and should consider potential infection with Coxiella burnetii. The positive rate of IgG antibody was 68% in the slaughterhouse workers.
ABSTRACT
Onosma dichroanthum Boiss. is one of the most important mountainous medicinal plants in Iran. This study was done to determine the biomechanical evaluation of root extract ointment of Onosma dichroanthum Boiss. on wound healing in rats. In this experimental study, 18 male adult Wistar rats were randomly allocated into control [I], vehicle [II] and treated group with ointment containing of extract 1% of root of Onosma dichroanthum Boiss, [III]. 20 mm vertical skin incision wound were made on rats back side. The assessment of the wound healing was carried out at day 14. At the end of study, rats were sacrificed, skin sample were extracted and evaluated by biomechanical method [maximum force, elastic stiffness, energy absorption]. There was no significant difference in biomechanical parameters among the treated, vehicle and control groups. Topical application of Onosma dichroanthum Boiss. root have no effect on healing of skin wound in animal model
Subject(s)
Wound Healing , Plant Extracts/pharmacology , Plant Roots , Plant Preparations , Plant Extracts , Models, Animal , Rats, Wistar , Plants, MedicinalABSTRACT
The Wnt/beta- The Wnt/beta-catenin signaling pathway is involved in many developmental processes in both fetal and adult life; its abnormalities can lead to disorders including several types of cancers and malfunction of specific cells and tissues in both animals and humans. Its role in reproductive processes has been proven. This study was designed to evaluate the expression of the key regulator of this signaling pathway GSK3-beta and its presumed role in azoospermia. WNT3[a] protein concentration and GSK3-beta gene expression levels were measured and compared between two groups of infertile men. The test groups consisted of 10 patients with obstructive and 10 non-obstructive azoospermia. The control group was selected among healthy men after vasectomies that were willing to conceive a child using a testicular biopsy technique. Samples were obtained by testicular biopsy and screened for the most common mutations [84, 86 and 255] in the SRY region before analyzing. GSK3-beta gene expression was assessed quantitatively by real time-PCR. The WNT3[a] protein concentration had no significant difference between the two test groups and controls. Expression of GSK3-beta was down-regulated in non-obstructive azoospermia [3.10 +/- 0.19] compared with normal [7.12 +/- 0.39] and obstructive azoospermia [6.32 +/- 0.42] groups [p=0.001]. Down-regulation of GSK-3beta may cause to non-obstructive azoospermia. Regulation and modification of GSK-3beta gene expression by drugs could be used as a therapeutic solution
ABSTRACT
The aim of this investigation was to improve the storage stability and survival rate of an intravesical BCG product, manufactured with an attenuated strain of Mycobacterium bovis [Pasteur strain 1173P2 of BCG] in the presence of sodium glutamate. Formulations with various concentrations of trehalose [a known protectant] were developed as liquid and lyophilized forms. Formulations were evaluated by different methods, including optical density measurement, safety assessment, skin reaction test, moisture content determination, viability assay, bacterial and fungal contaminations and the results were compared with those obtained for sodium glutamate-containing formulations. The stability tests were also carried out in various storage durations and different temperatures. To develop the lyophilization protocol, glass transition temperatures in the presence of both stabilizers were determined using differential scanning calorimetry. In general, results showed that trehalose could considerably increase the stability of the product against freezing and drying processes, increase the survival rate even in the liquid formulations, as well as the production of an acceptable cake. However, further studies are required to optimize the product characteristics
ABSTRACT
Tubal ectopic pregnancy [tEP] is the most common type of extra-uterine pregnancy and the most common cause of maternal mortality. Nitric oxide [NO] is a molecule that incorporates in many physiological processes of female reproductive system. Recent studies have demonstrated the possible role of endothelial isoform of nitric oxide synthase [eNOS] enzyme in the regulation of many reproductive events that occur in the fallopian tube [FT]. The aim of this study was to evaluate the expression of eNOS in the FTs of women with tEP. In this case-control study, a total number of 30FTs samples were obtained from three groups including: 10 FTs of women that bearing an EP, 10 FTs from the non-pregnant women at luteal phase of the menstrual cycle, and 10 FTs of healthy pregnant women [n=10]. Samples were fixed in 10% buffered formalin and then were evaluated by immunohistochemistry. Localization of eNOS was seen in secretory and ciliated luminal epithelium and vascular endothelium of all groups. However, we did not observed the expression of eNOS in smooth muscle cells of all groups. Expression of eNOS in luminal epithelium of women with EP compared to non-pregnant women at luteal phase of menstrual cycle and healthy pregnant group showed statistically significant increase [p=0.00]. Significant difference in expression of eNOS was not observed in luminal epithelium of FTs of women at luteal phase compared to healthy pregnant groups [p=0.78]. This study indicates that changes in expression of eNOS in luminal epithelium of FT may lead to development of EP.
ABSTRACT
<p><b>OBJECTIVE</b>To determine the presence of antibodies against phase II among slaughterhouse workers in Kerman, southeast of Iran.</p><p><b>METHODS</b>The antibody titers of the serum samples were measured by enzyme-linked immuno sorbent assay using phase II Coxiella burnetii as the antigen [kit (Virion\Serion, Wurzburg, Germany) according to the manufacturer's protocol].</p><p><b>RESULTS</b>The positive rate of IgG antibody was 68% in the slaughterhouse workers.</p><p><b>CONCLUSIONS</b>Our findings suggest that slaughterhouse workers in Kerman area have a higher risk of infection and should consider potential infection with Coxiella burnetii.</p>
ABSTRACT
Penicillin G acylase was immobilized onto iron oxide nanoparticles coated with polyethyleneimine and then cross linked with glutaraldehyde solution. The FTIR spectrum of immobilized enzyme showed peak at 1648cm-1 which can be attributed to the C=N bonds of Schiff’s base linkage formed between glutaraldehyde and amino group of penicillin G acylase. By considering the FTIR spectrum of nano particle coated with polyethyleneimine, adsorption of penicillin G acylase has taken place and then glutaraldehyde cross linked enzyme onto activated support. Catalytic properties of nano penicillin G acylase were improved upon immobilization as compared to its free counterpart. The optimal pH and temperature were determined to be 7.0, 10.0, 50 and 75ºC for free and immobilized penicillin G acylase, respectively. Thermal stabilities of both nano and free penicillin G acylase were studied .The Km value of immobilized nanozyme was calculated from Lineweaver Burck plot to be 0.23 μM while that of free penicillin G acylase was 0.28μM. In this way nano penicillin G acylase with improved catalytic properties was developed as compared to its soluble counterpart.
ABSTRACT
Recently, milk has been suggested as one of the main sources of iodine for human intake. However, there is the potential for problems resulting from excessive amounts of iodine in cow milk. Supplementary feed iodine increases milk iodine and is apparently the cause of many of the high milk iodine values. Iodine teat dips and sanitizers also contribute to increased milk iodine values. Variability of these practices may result in variation of iodine concentrations in milk. In addition, the transportation and processing phases of milk production, where iodophor sanitizers are used, season, geographic region, stage of lactation and iodine antagonists in feeds, may influence milk iodine concentration. It seems that Iodine concentration in milk should be monitored, regularly.
ABSTRACT
Purpose: The aim of this study was to assess the production of recombinant cholera toxin B subunit (rCTB) protein in two different expression systems (pAE_ctxB and pQE_ctxB constructs) in Escherichia coli BL21 (DE3). Materials and Methods: The ctxB fragment was amplified from Vibrio cholerae O 1 ATCC14035 and cloned in pGETM-T easy vector after which it was transformed to E. coli Top 10F' and grown on LB-ampicillin agar medium. Sequence analysis confirmed the complete ctxB gene sequence in the construct which was further subcloned to pQE-30 vector. The construct was subsequently transformed to E. coli M15 (pREP4). The recombinant pAE_ctxB and pQE_ctxB were transformed to competent E. coli BL21 (DE3) cells to express CTB protein. Result: Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis showed the maximum expression of rCTB in both systems at 5 h after induction and western blot analysis confirmed the presence of recombinant CTB in blotting membranes. Conclusion: Expression of rCTB in pAE_ctxB construct was more efficient (15-fold) than pQE_ctxB, and it seems that Lac UV5 in E. coli BL21 (DE3) is more compatible with the former construct. This expression system can be used to produce recombinant CTB in high yield which may enable us to study the oral tolerance or mucosal adjuvant properties of rCTB using animal models.
ABSTRACT
Although there is convincing data in support of the effectiveness of hyperthermia in tumor therapy, the molecular mechanisms underlying the clinical effects of hyperthermia are still poorly understood. To investigate natural killer [NK] cell cytotoxicity against heat-treated SW-872 and HeLa tumor cell lines. NKG2D ligands and HLA class I transcription were examined using quantitative real-time PCR in treated tumor cell lines at 0, 2, 4, 6 and 12 h following thermal treatment at 39 Degree °C and 42 Degree °C for 1 h. The expression of MICA/B, ULBP1 and ULBP2 were also determined by flow cytometry. NK92-MI cytotoxic activity against heat-treated target cell lines was assessed by LDH release as well as annexin-V and 7-AAD assays. Our results showed that heat treatment at 39°C improved the cytolytic activity of NK cells against SW-872 cells without increasing NKG2D ligand concentration or decreasing HLA class I levels. The observed increase in the cytotoxicity of NK cells against SW-872 cells after hyperthermia does not coincide with changes in MICA/B, ULBP1 and ULBP2 ligands of NKG2, however, the expression of other ligands in target cells may have made the cells susceptible to the cytotoxic effect of NK cells
ABSTRACT
Therapeutic effects of various treatment options in wound healing have been one of the most controversial issues in surgical science. The present study was carried out to examine and compare the effects of Aloe vera gel, thyroid hormone cream and silver sulfadiazine cream onsutured incisions in Wistar rats. In a randomized controlled trial, thirty-six Wistar male rats, 250 to 300 g, received surgical incisions followed by topical application of Aloe vera gel, thyroid hormone cream and silver sulfadiazine 1%. To assess the efficacy of each treatment technique, a histological approach was used to evaluate the mean number of fibroblasts, macrophages, neutrophils, blood vessel sections and thickness of the regenerating epithelium and dermis on days 4, 7 and 14. Re-epithelialization and angiogenesis were significantly improved in Aloe vera gel group compared with the other treatments while thyroid hormone cream had positive effects on day 4 (P< or =0.05). Topical administration of Aloe vera gel is recommended as the treatment of choice for surgical incisions.
Subject(s)
Animals , Humans , Male , Rats , Administration, Topical , Aloe , Blood Vessels , Dermis , Epithelium , Fibroblasts , Glycosaminoglycans , Macrophages , Neutrophils , Rats, Wistar , Re-Epithelialization , Silver Sulfadiazine , Thyroid Gland , Thyroid Hormones , Wound HealingABSTRACT
Many research studies report the healing effects of Aloe Vera, thyroid hormone cream and silver sulfadiazine. However, the effects of these therapeutic agents are not well understood and have not been compared in one study. This study aimed at investigating the effects of topical application of an Aloe vera gel, a thyroid hormone cream and a silver sulfadiazine cream on the healing of skin wounds surgically induced in Wistar rats for determining the treatment of choice. In a randomized controlled trial, twelve male rats, aged 120 days and with a mean weight of 250 to 300 g, were divided randomly into 5 groups based on drug treatments: Aloe vera gel (AV), thyroid hormone cream (TC), silver sulfadiazine 1% (S), vehicle (V) and control. To evaluate the efficacy of each treatment technique, a biomechanical approach was used to assess tensile stress after 14 days of treatment. Tensile stress was significantly improved in the Aloe vera gel group as compared with the other four groups (P< or =0.05). While the other treatment options resulted in better healing than the control group, this difference was not significant. We conclude that Aloe vera topical application accelerated the healing process more than thyroid hormone, silver sulfadiazine and vehicle in surgically induced incisions in rats.
Subject(s)
Aged , Animals , Humans , Male , Rats , Aloe , Rats, Wistar , Silver , Silver Sulfadiazine , Skin , Thyroid Gland , Wound HealingABSTRACT
The aim of this study was to evaluate the effects of dexamethasone on striatal dopaminergic, glutamatergic and gamma amino butyric acid [GABA] ergic neurotransmission in normal and parkinsonian rats. Dexamethasone [0.15, 0.30, 0.60 and 0.8 mg/kg] was administered to normal or parkinsonian rats [i.p.] followed by the analysis of the striatal neurotransmitters concentrations. Additionally, the effect of dexamethasone on the damaged Substantian nigra pars compata [SNc] neurons has been investigated. Dexamethasone resulted in decreased level of striatum glutamatergic-GABAergic and enhanced dopaminergic neurotransmission in normal and parkinsonian rats. In addition, acute treatment with dexamethasone did not improve the lesion at all. These findings suggest the new therapeutic mechanism of action for dexamethasone in Parkinson's disease animal model
Subject(s)
Male , Animals, Laboratory , Dexamethasone/pharmacology , Rats, Wistar , Models, Animal , Parkinson Disease/veterinary , Parkinson Disease/therapy , Dopamine , gamma-Aminobutyric Acid , Glutamic AcidABSTRACT
The attempts were made to describe the development of a whole cell immobilization of P. pastoris by entrapping the cells in polyacrylamide gel beads. The alcohol oxidase activity of the whole cell Pichia pastoris was evaluated in comparison with yeast biomass production. Methylotrophic yeast P. pastoris was obtained from Collection of Standard Microorganisms, Department of Bacterial Vaccines, Pasteur Institute of Iran [CSMPI]. Stock culture was maintained on YPD agar plates. Alcohol oxidase was strongly induced by addition of 0.5% methanol as the carbon source. The cells were harvested by centrifugation then permeabilized. Finally the cells were immobilized in polyacrylamide gel beads. The activity of alcohol oxidase was determined by method of Tane et al. At the end of the logarithmic phase of cell culture, the alcohol oxidase activity of the whole cell P. Pastoris reached the highest level. In comparison, the alcohol oxidase activity was measured in an immobilized P. pastoris when entrapped in polyacrylamide gel beads. The alcohol oxidase activity of cells was induced by addition of 0.5% methanol as the carbon source. The cells were permeabilized by cetyltrimethylammonium bromide [CTAB] and immobilized. CTAB was also found to increase the gel permeability. Alcohol oxidase activity of immobilized cells was then quantitated by ABTS/POD spectrophotometric method at OD[420]. There was a 14% increase in alcohol oxidase activity in immobilized cells as compared with free cells. By addition of 2-butanol as a substrate, the relative activity of alcohol oxidase was significantly higher as compared with other substrates added to the reaction media. Immobilization of cells could eliminate lengthy and expensive procedures of enzyme separation and purification, protect and stabilize enzyme activity, and perform easy separation of the enzyme from the reaction media
Subject(s)
Oxidation-ReductionABSTRACT
The aim of present investigation was to determine the effects of low-level He-Ne laser therapy on biomechanical property of skin wound of healthy and streptozotocin induced diabetic [STZ-D] rats. The study was performed by experimental method. 36 male adult Wistar rats weighing above 250 gr. were used. Rats were divided equally into control and experimental groups, each group [n=18] were equally divided into 3 subgroups in order to radiate 3 different energy densities of laser. Weight of healthy and diabetic rats were recorded in the beginning and at the end of study. Blood glucose of rats in the beginning of study was recorded and rats with more than 120 mg/dl were excluded from study. Diabetes was induced by one time intra peritoneal injection of 55 mg/kg STZ. After one month, hyperglycemia was established in experimental group. Two 15-mm, vertical incision wounds were made on the dorsum of rats. Three groups of healthy and diabetic rats were received 22.4J/cm[2], 1.2J/cm[2] and 4J/cm[2] energy densities He-Ne laser for two weeks. At the end of study, rats were killed and skin sample were extracted and were submitted to a biomechanical evaluation [maximum force] examination. Data was analyzed by paired student t test methods. Mean value of blood glucose of diabetic rats was 518.37 +/- 23.3. Laser-treated healthy rats with 1.2J/cm[2] energy density showed significant increase of maximum force [p=0.05]. Laser-treated diabetic rats with 4J/cm[2] energy density showed significant increase of maximum force [p=0.05]. It seems ideal parameters for effectiveness of Low- Level He-Ne laser in healthy and diabetic rats are different. Wounds of diabetic rats should be radiated with more energy density of low- level laser for accelerating wound healing process in comparison with healthy rats
Subject(s)
Animals, Laboratory , Diabetes Mellitus, Experimental , Streptozocin , Lasers, Gas , Low-Level Light Therapy , RatsABSTRACT
Background: brucellosis is one of the five common bacterial zoonosis caused by a gram negative, non-spore forming, and facultative intracellular bacterial organism belonging to the genus Brucella. Although brucellosis is considered as a health problem for both men and domestic animals in many countries, any licensed human vaccine has not been designed and produced for it yet. To overcome the problem, currently, antigenic determinants of Brucella cell wall e.g. outer membrane proteins [OMPs] and lipopolysaccharide [LPS] are considered as potential candidates to develop subunit vaccines
Materials and Methods: brucella abortus S99 used in the present study is obtained from the standard bacterial collection of Institute Pasteur of Iran. OMPs were extracted by deoxy cholate extraction technique and further purification performed by sequential centrifugation and ultracentrifugation. Protein concentration was determined using the Nano drop ND-10000 spectrophotometer. SDS-polyacrylamide gel electrophoresis [SDS- PAGE] was performed to determine the electrophoretic pattern and the molecular weight of the extracted OMP samples
Results: OMPs concentration of B.abortus S99 has been measured and reported as 6.27 mg/ml. SDS-PAGE analysis indicated one protein band in the range of 36-38 kDa which would be classified as the porins of B.abortus S99
Conclusion: extraction of B.abortus S99 OMPs with the applied method in the present study produced a satisfactory yield of OMPs. These proteins belonging to the second group of OMPs, called porins