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Effects of Cadmium Poisoning on the Expression of NF-kB/MAPK and CD163 Genes in Rat Adrenal Gland Texto completo
2023
Gholami, Atieh | Marjani, Zahra | Hesaraki, Saeed
BACKGROUND: Cadmium is a heavy metal harmful to animals and humans. Exposure to it causes inflammation, apoptosis, or necrosis in numerous tissues, including the adrenal.OBJECTIVES: The present research investigates the effect of cadmium toxicity on the expression of genes involved in inflammation and fibrosis. Inflammation increases the rate of parenchymal cell death, and fibrosis will only fill the place of dead cells without being able to perform the function of the primary parenchyma.METHODS: In this research, cadmium chloride with a concentration of 20 mg/kg was added to the diet of ten mice in two groups of five. On the 30th day of the study, the adrenal glands were quickly sent to the laboratory. The expression of NF-kB/MAPK, hematoxylin, eosin tissue staining, and immunohistochemistry (CD163) were performed.RESULTS: The inflammation mentioned in others’ research can also be associated with the activation of the nuclear factor kappa (NF-kB) pathway. NF-κB gene products initiate mitogen-activated protein kinase (MAPK) and p38 pathways. Previous studies indicate that MAPK induces necrosis or apoptosis in tissues. In histopathology, dense and possibly pyknosis nuclei are more common in the cadmium group. The higher expression of the CD163 molecule in the cadmium group reveals the beginning of the fibrosis process after chronic inflammation.CONCLUSIONS: This report provides more basic data to investigate the mechanism of adrenal damage in cadmium poisoning. Cadmium causes the death of cells by affecting the inflammatory pathways. Additionally, the stimulation of the fibrosis process causes greater irreparable damage to the damaged tissue of the adrenal gland.
Mostrar más [+] Menos [-]Anatomic and Excretory Urography Investigation of the Kidneys, Ureters and Urinary Bladder in Guinea Pig Texto completo
2023
Azimzadeh, Saba | Alizadeh, Siamak | Hosseinchi, Mohammad Reza
BACKGROUND: Excretory urography is a method of imaging the kidneys, ureters, and urinary bladder which uses contrast medium containing the iodine compounds.OBJECTIVES: This study aimed to evaluate the anatomical structures of urinary tract in the nephrogram, pilogram, and cystogram phases, and determine the exact standard for the size of kidneys, ureters and urinary bladder in guinea pigs to be used to interpret the results, and clinical decisions.METHODS: This study was carried out on 10 guinea pigs with a mean age of 12±1.33 months and average weight of 1.12±0.18 kg. Before to the administration of contrast medium, each guinea pig was fast and Dimethicone 20 mg/kg was given orally. At the time of administration of contrast agent, each animal was sedated by using Ketamine 30 mg/kg and diazepam 5 mg/kg cocktail, and then 1500 mgI/kg of meglumine compound 60 % was injected subcutaneously over the shoulder area. Ventrodorsales and lateral abdominal X-rays were taken, thereafter every 5 minutes up to 60 minutes to complete the pylogram phase. In lateral radiographs of each guinea pig, the length of the body of the second lumbar vertebra was measured to be used as an indicator in determining the standard size of the kidneys.RESULTS: Based on the results of this study, the average length, width, and thickness of the right kidney compared to the length of the second lumbar vertebra were 2.19, 1.64, and 1.33 cm, and in the left kidney of 2.09, 1.53, and 1.41 cm and this average in right and left ureter was 6.41 and 6.22 cm, respectively.CONCLUSIONS: The exact standards can be used in the interpretation of results, and clinical decisions to determine the normal and abnormal size of kidneys, ureters and bladder in the guinea pigs.
Mostrar más [+] Menos [-]A Preliminary Investigation of the Haemoproteus Infection in Domestic Pigeons of Torkaman County, Iran by Microscopic and Molecular Methods Texto completo
2023
Iri, Saeid | Firouzivand, Yaghoub | Hosseinzadeh, Somayyeh
BACKGROUND: The protozoan Haemoproteus belongs to the Phylum Apicomplexa, Class Sporozoa, and Order Haemosporina. Avian haemosporidian are protozoan parasites that use birds as hosts around the world. Many species of wild and domestic doves are natural hosts of different species of Haemoproteus. Blood-sucking arthropods are the main vectors of these blood parasites.OBJECTIVES: The aim of this study was the microscopic and molecular investigation of the protozoan Haemoproteus columbae in the blood of infected pigeons in Torkaman County, Iran.METHODS: Blood samples and tubes containing ethylenediaminetetraacetic acid (EDTA) anticoagulant were collected from 96 domestic pigeons randomly from 14 pigeon lofts and different parts of Torkaman County.Pigeons were also inspected for infection with the host-vector Pseudolynchia canariensis. In the next step, blood smears were stained with Giemsa and examined microscopically. Also, blood tubes containing EDTA were tested by PCR method on the cytochrome b gene.RESULTS: Microscopic and molecular examination of peripheral blood showed that 62 (64.58 %) and 73 (76.04 %) of the investigated pigeons were contaminated, respectively. Of the 62 infected pigeons infected with the Haemoproteus, 28 pigeons (66.66 %) were male, and 34 (62.96 %) were female. Also, the infestation with Pseudolynchia canariensis was observed in 4 (28.57 %) pigeon lofts.CONCLUSIONS: The preliminary investigation shows the high rate of Haemoproteus infection in pigeons in Torkaman County. Further studies to determine the prevalence and accurate identification of the species infecting pigeons in this region require PCR testing and sequencing of infected blood samples.
Mostrar más [+] Menos [-]The Effect of Curcumin on the Structure of Mouse Ovary After Treatment With Goserelin and Cyclophosphamide Texto completo
2023
Azarmi, Sareh | Talebkhan Garoussi, Massoud | Tajik, Parviz | Hosseini Pajooh, Khosro | Sasani, Farhang | Jahanroshan, Navid
BACKGROUND: Protection from reproductive damage is essential in chemotherapy medicines for cancer patients.OBJECTIVES: This study aims to examine the effect of curcumin on the structure of the ovary of mice after treatment with goserelin and cyclophosphamide.METHODS: One hundred and ten BALB/C mice with 3 regular consecutive periods of the estrous cycle were divided into 11 groups of 10 each. No medicine was used in the control group. The treatment groups were as follows: 1) cyclophosphamide, 2 to 5) cyclophosphamide with curcumin with a dose of 100, 200, 300, and 400 mg/kg, respectively, 6) goserelin, 7 to 10) goserelin together with curcumin with a dose 100, 200, 300, 400 mg/kg, respectively. The luteinizing hormone (LH) and follicle-stimulating hormone (FSH) of serums were evaluated using ELISA. Morphologic and morphometric of ovaries were assessed.RESULTS: The total number of follicles, primary, secondary, periantral, and antral follicles, in the goserelin and cyclophosphamide group, was significantly reduced compared with the control group (P<0.05). Cyclophosphamide and goserelin with different doses of curcumin showed a significant increase in the total number of follicles, primary, periantral, and antral follicles compared to the group treated with cyclophosphamide and goserelin alone (P<0.05). Curcumin (200, 300, and 400 mg/kg) and cyclophosphamide, compared to the cyclophosphamide group, significantly increased the quality of zona pellucida (P<0.05). Cyclophosphamide and goserelin caused a significant decrease in FSH and LH (P<0.05). Cyclophosphamide with different doses of curcumin showed a significant increase in LH compared to the group treated with cyclophosphamide alone (P<0.05). Goserelin with a 400 mg/kg curcumin dose significantly increased LH compared to goserelin alone (P<0.05). The amount of FSH in the cyclophosphamide groups with curcumin increased compared considerably to cyclophosphamide alone (P<0.05). The groups of goserelin with curcumin showed a significant increase in FSH compared to those of goserelin alone (P<0.05).CONCLUSIONS: Curcumin can protect the reproductive system of mice from the damage caused by the administration of cyclophosphamide and goserelin.
Mostrar más [+] Menos [-]Evaluating PCR-RFLP Technique in Identifying Genetic Diversity Clostridium perfringens Biotype A Texto completo
2023
Mosahasankhani, Hamid | Shamsaddini Bafti, Mehrdad | Kazemipour, Nadia | Alimolaei, Mojtaba | Rokhbakhsh-Zamin, Farokh
BACKGROUND: Clostridium perfringens (C. perfringens) is an anaerobic Gram-positive bacillus with spores, whose biotype A is responsible for a variety of diseases, including intestinal inflammation, bloody diarrhea, and gas gangrene, and hemorrhagic bowel syndrome. Genetic variety can explain the bacteria’s phenotypic diversity, geographic distribution, host specificity, pathogenicity, antibiotic resistance, and virulence. A molecular method using the pattern of DNA bands classifies bacteria based on the size of fragments produced by enzymatic digestion of the genome.OBJECTIVES: This study aims to standardize the polymerase chain reaction (PCR)- restriction fragment length polymorphism (RFLP) method in identifying the genetic diversity of C. perfringens biotype A isolates.METHODS: The genomic DNA of the investigated strains was extracted, and the complete sequence of the alpha toxin gene locus was synthesized using specific primers designed by PCR technique. Enzymatic cleavage of the synthesized amplicons was performed with the Mse l restriction enzyme, and the resulting fragments were separated by electrophoresis and analyzed by ImageJ and NTSYSPC software.RESULTS: The findings showed that the alpha toxin gene locus sequence may change and is not conserved. In this research, 4 different patterns were identified based on enzymatic cleavage. Mutations in this locus can lead to diversity in C. perfringens biotype A and the creation of new strains.CONCLUSIONS: The results of this research showed that the alpha toxin gene locus could be considered a DNA molecular marker in C. perfringens, and the PCR-RFLP technique can be used as a tool for typing this bacterium and estimating the phylogenetic relationships through comparative studies of nucleotide sequences.
Mostrar más [+] Menos [-]Evaluation of Skin Repairing and Antifungal Properties of Alcoholic Extract of Laleh abbasi (Mirabilis jalapa) Leaf on Induced Wounds in Laboratory White Rat Model Texto completo
2023
Zamani Raad, Behran | Mardjanmehr, Seyed Hossein | Sasani, Farhang | Khosravi, Alireza | Gharagozlou, Mohammad Javad
BACKGROUND: Based on the historical and recently published documents, it has been demonstrated that Mirabilis jalapa as a herbal medicine may be used as remedies for various health problems included wound healing purposes.OBJECTIVES: The present study aimed to investigate the effect of ethanolic extract of Laleh abbasi green leaf on healing open wounds induced by skin puncture in the back of rats.METHODS: Collecting and drying Laleh abbasi leaves, leaf extracting through Soxhlet procedure, analyzing the extract via gc / ms method, and preparing eucerin-based extract ointment were done according to recommended routine procedures. Herein, we recruited 40 male rats that were randomly divided into five groups of eight, namely the control, phenytoin treatment, eucerin, 5% eucerin extract, and 7.5% eucerin extract ointment treatment groups. Skin puncture and application of ointments on the induced wounds were carried out. Subsequently, tissue samples were taken on days 3, 7, 10, and 14, followed by which histological slides were prepared and stained with H&E and Masson’s trichrome staining methods. In vitro mycological studies were conducted using opportunistic fungi, including Candida, Mucor, and Aspergillus species.RESULTS: Based on the macroscopic evaluations of the wound healing process and microscopic assessments of tissue samples stained with Harris H&E and Masson’s trichrome methods, the groups treated with eucerin-based ointments containing ethanolic extract of Laleh abbasi leaf had statistically significant positive wound healing responses compared to the other groups. However, the 7.5% ointment group showed statistically better responses compared to the 5% ointment group. The data obtained in the present preliminary experiment on rat model indicated that the extract could facilitate the wound healing process in terms of healing parameters, such as accelerating epithelium repair, creating a favorable inflammatory reaction, angiogenesis, fibroplasia, and collagen precipitation. The antifungal effects of ethanolic extract of Laleh abbasi leaves on Aspergillus fumigatos, fusarium, Candida albicans and Candida cruzei were demonstrated in vitro using saboro dextrose agar medium.CONCLUSIONS: According to the findings of this experimental study and the findings of other researchers, it can be concluded that ethanolic extract from Laleh abbasi leaves is of healing and antifungal properties.
Mostrar más [+] Menos [-]Comparison to Methods; Serum Antibody ELISA and Fecal Nested-PCR to Diagnose Mycobacterium avium Subspecies Paratuberculosis Subspecies Infection in Cattle Texto completo
2023
Kolivand, Ali | Haji Hajikolaei, Mohammad Rahim | Nouri, Mohammad | Khosravi, Mohammad | Gharibi, Dariush
BACKGROUND: Mycobacterium avium subspecies paratuberculosis is the cause of a common disease in dairy herds. Early diagnosis of paratuberculosis infection can improve Johne’s disease control programs.OBJECTIVES: This study aimed to compare the sensitivity, and specificity to methods; blood serum ELISA and stool Nested-PCR for the detection of Mycobacterium avium subspecies paratuberculosis infection in dairy cattle.METHODS: A commercial ELISA kit was used to perform the absorbed ELISA test, which was conducted after exposing serum samples to Mycobacterium phlei antigens to limit cross-reactions. Nested-PCR test was performed using nucleotide sequences related to specific MAP gene fragments, i.e. IS900.RESULTS: As a result of the ELISA antibodies kit, out of the total 2203 serum samples, 112 samples were positive (5.08 %) and 2091 samples were negative (94.92 %). The results of Nested-PCR tests of rectal feces showed that out of 59 cows with the positive results in serum ELISA, 47 (79.66 %) samples were positive and 12 (20.34 %) samples were negative. Moreover, out of 31 cattle with a negative result on the ELISA test, 15 (48.38%) and 16 cattle (51.62 %) had positive and negative results, respectively, on the nested PCR tests of the feces samples.CONCLUSIONS: Due to the low sensitivity of PCR compared to ELISA, the positive and negative predictive values, and the accuracy of ELISA test, as well as the high cost and time-consuming nature of PCR and the need for more and more complex facilities than ELISA, the authors concluded that ELISA is a more suitable method for screening and epidemiological studies than PCR.
Mostrar más [+] Menos [-]Evaluation of the Antibacterial Effect of Carvacrol Alone and in Combination with the Antibiotic Cefixime Against Escherichia coli O157:H7 Texto completo
2023
Asadi, Sepideh | Nayeri Fasaei, Bahar | Zahraei Salehi, Taghi | Yahya Rayat, Ramak | Shams, Nemat
BACKGROUND: The use of plant compounds and their derivatives, such as extracts and essential oils, for combating infectious agents has attracted a great deal of scientific attention. One of the active antimicrobial compounds with plant origin is carvacrol.OBJECTIVES: The present study aimed to evaluate the antibacterial activity of carvacrol alone and in combination with the antibiotic cefixime against Escherichia coli O157:H7.METHODS: The antibacterial properties of carvacrol and cefexime were evaluated by determining the Minimum Inhibitory Concentration (MIC), Minimum Bactericidal Concentration (MBC), and disk diffusion method. The checkerboard assay was used to evaluate the interaction between the carvacrol and cefexime and to determine the fractional inhibitory concentration.RESULTS: The results showed that the MIC and MBC of carvacrol and cefexime against E.coli O157:H7 were 250, 250 μg/ml (MIC, MBC) and 128, 128 μg/ml (MIC, MBC), respectively. In the checkerboard test, carvacrol was found to have a synergistic interaction with antibiotic cefixime against E. coli O157:H7 (FIC index=0.5).CONCLUSIONS: Due to the significant antibacterial activity of carvacrol, the present study introduced this agent as a new antibacterial drug with a natural origin. In addition, since carvacrol significantly increased the antibacterial potential of cefixime (synergistic properties), it could be considered as an effective compound for increasing the antibacterial power of cefixime antibiotic.
Mostrar más [+] Menos [-]Tracking and Identifying Enterobacteriaceae Contamination in Darkling Beetles (Tenebrionidae) as One of the Reservoirs of Bacteria Persistence Poultry Farms Texto completo
2023
Mokhtari, Pegah | Jalalizand, Alireza | Mahmoudi, Esmaeil | Ghalamkari, Gholamreza
BACKGROUND: Poultry farming is one of the most productive and economic agricultural sectors. However, the bacterial contamination and the activity of darkling beetles (Tenebrionidae) as a potential reservoir of Salmonella in meat poultry farms can inflict direct and indirect damages.OBJECTIVES: The present study aimed to identify the darkling beetles and their accompanying Enterobacteriaceae contamination in Isfahan chicken farms.METHODS: Darkling beetles were collected and identified based on their morphological aspects from different parts of 16 poultry farms (4 from each geographical area) in Isfahan Province, Iran. Then, 80 samples of darkling beetles were cultured on selective-differential media culture of the Enterobacteriaceae family using the homogenization and enrichment method. The isolated bacteria were identified based on physiological and molecular characteristics. Also, specific antisera were used to determine serological groups.RESULTS: The results revealed that all collected darkling beetles’ samples belonged to the species Alphitobius diaperinus (Col., Tenebrionidae), and from 80 microbial culture samples from the beetles, isolated bacteria belonged into 4 genera: Escherichia sp. (20 isolates, 25 %), Klebsiella sp. (8 isolates, 10 %), Proteus sp. (22 isolates, 27.5 %), and Salmonella sp. (30 isolates, 37.5 %). Among them, the Salmonella genus accounted for the highest percentage of darkling beetles’ contamination. In the serological assay, the isolated Salmonella were classified into two serogroups, A (23 isolates, 76.67 %) and C (C2 and C3) (7 isolates, 23.33 %), which the A serogroup was the most frequent.CONCLUSIONS: In this study, the A. diaperinus species was isolated and identified for the first time from poultry farms, and this pest, with a high percentage of Salmonella infection, is introduced as one of the reservoir sources of bacterial contamination in the broiler farms.
Mostrar más [+] Menos [-]Evaluating the Effect of Culture Supernatant of Pseudomonas aeruginosa on Removing the Inhibitory Effect of Heparin in Real-Time PCR Test Texto completo
2023
Ashrafi, Aysan | Staji, Hamid | Keramati, Keyvan
BACKGROUND: Heparin is a sulfated glycosaminoglycan. Blood is a common source for DNA detection in all kinds of samples, and anticoagulants such as heparin and ethylenediaminetetraacetic acid (EDTA) are used to prevent coagulation. Because heparin has a strong inhibitory effect on polymerase chain reaction (PCR), it is not used in samples that will be tracked by DNA. There are physical, chemical, and enzymatic methods to eliminate the inhibitory effect of heparin on PCR test.OBJECTIVES: First, to compare the intensity of the inhibitory effect of two anticoagulants, heparin, and EDTA, on the Real-Time PCR (qPCR), and then to investigate the impact of the heparinase enzyme present in the medium culture extract of Pseudomonas aeruginosa, on removing the inhibitory effect of heparin during the real-time PCR.METHODS: In the present study, two blood samples containing heparin and EDTA were subjected to a real-time PCR test to check the intensity of the inhibitory effect. Then, the medium culture extract of Pseudomonas aeruginosa was added to the heparinized blood sample infected with Escherichia coli bacteria in two groups with different conditions. In the first group, the DNA in the heparinized blood sample was extracted by the phenol-chloroform isoamyl alcohol method. Then, these samples were incubated with the extract of Pseudomonas aeruginosa bacteria culture medium at different hours, but in the second group, the samples were incubated at different hours before DNA extraction. Also, the DNA concentration in both groups was measured by a Nanodrop device, and finally, all samples were subjected to a real-time PCR test.RESULTS: The results of the research samples showed that although the heparinized blood sample contains more DNA concentration than the EDTA blood sample, it completely prevents genome replication. Also, incubating heparinized blood with Pseudomonas aeruginosa culture medium extract before DNA extraction for more than 24 hours removes the inhibitory effect of heparin during the real-time PCR, even at a lower cycle threshold than the EDTA-containing sample.CONCLUSIONS: The Pseudomonas aeruginosa culture medium extract may enable researchers to use heparinized blood samples for genome amplification and diagnosis without using expensive and limited commercial heparinase enzyme.
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