ABSTRACT
Purpose@#Newcastle disease (ND) represents a major viral disease across the world which imposes high costs to poultry producers for vaccination. Hemagglutinin-neuraminidase (HN) and fusion (F) proteins are the major immunogenic epitopes of Newcastle disease virus and hence, have been the main targets for development of anti-ND vaccines. This paper reports transient expression of a synthetic gene composing of four tandem repeats of HN and three tandem repeats of F epitopes in maize leaves as initial step toward production of recombinant vaccine against ND. @*Materials and Methods@#The synthetic gene was cloned in pBI121 plasmid to yield an expression vector. The vector was sophisticated by the addition of AUG codon, polyhistidine-tag, tobacco mosaic virus omega sequence, stop codon, and restriction sites. Leaf transformation was conducted by the agroinfiltration method. Molecular detection assays including polymerase chain reaction, reverse transcription-polymerase chain reaction and enzyme-linked immunosorbent assay (ELISA) were carried out to evaluate transgene expression in infiltrated leaves of the corn plant. @*Results@#The result obtained in this research revealed that the transgene was transcribed and translated in maize leaves only 48 hours after infiltration. In the second phase of the experiment, the expressed protein was injected into rabbits. The result of the ELISA assay indicated induction of immune response in the rabbits after injection with the heterologous protein. @*Conclusion@#These results confirm the feasibility of agroinfiltration for transient gene expression of viral epitopes in monocot plants which naturally resist stable transformation by Agrobacterium tumefaciens. Practical implications of this finding are discussed in detail and some recommendations for future studies are proposed.
ABSTRACT
RFamide-related peptide-3 [RFRP-3] inhibits gonadotropin releasing hormone [GnRH] and luteinizing hormone [LH] secretion in rats. This study evaluates the effects of litter size and suckling intensity on RFRP mRNA expression in the dorsomedial hypothalamic nucleus [DMH] of rats. A total of 32 pregnant and 4 non-lactating ovariectomized [control group] Sprague-Dawley rats were used in this experimental study. Lactating rats were allotted to 8 equal groups. In 3 groups, the Utter size was adjusted to 5, 10, or 15 pups upon parturition. Dams were allowed to suckle their pups continuously until 8 days postpartum. In the other 3 groups, the litter size was adjusted to 5 pups following birth. These pups were separated from the dams for 6 hours on day 8 postpartum, after which the pups were allowed to suckle for 2.5, 5, or 7.5 minutes prior to killing the dams. In 2 groups, lactating rats with 10 and 15 pups were separated from their pups for 6 hours on day 8 postpartum. In these groups, the pups were allowed to suckle their dams for 5 minutes before the dams were killed. All rats were killed on day 8 postpartum and the DMH was removed from each rat. We evaluated RFRP mRNA expression using real-time polymerase chain reaction [PCR. The expression of RFRP mRNA in the DMH increased with increased litter size and suckling intensity compared to the controls. The effect of suckling intensity on the expression of RFRP mRNA was more pronounced compared to the litter size. Increased litter size and suckling intensity stimulated RFRP mRNA expression in the DMH which might contribute to lactation anestrus in rats
ABSTRACT
RFamide-related peptide-3 [RFRP-3] and kisspeptin [KiSS-1] are known to respectively inhibit and stimulate gonadotropin releasing hormone [GnRH] and luteinizing hormone [LH] secretion in rat. The aim of the present study was to evaluate the relative mRNA expression of RFRP-3 and KiSS-1 in the hypothalamus of pregnant rats. In a randomized controlled experimental study, the exact pregnancy day of 18 Sprague-Dawley rats were confirmed using the vaginal smear method and were equally assigned to three groups of days 7, 14 and 21 of pregnancy. Four non-pregnant female rats were ovariectomized and assigned as the control group. All rats were decapitated, and the dorsomedial hypothalamic nucleus [DMH] and the arcuate nucleus [ARC] for detection of KiSS-1 mRNA were separated from their hypothalamus to detect RFRP-3 and KiSS-1 mRNA respectively. Then, their relative expressions were compared between control and pregnant groups using real-time polymerase chain reaction [PCR]. The relative expression of RFRP-3 mRNA in DMH did not change significantly during pregnancy [p>0.01]. However, the relative expression of KiSS-1 mRNA in ARC was at its highest in day 7 of pregnancy and decreased until day 21 of pregnancy [p<0.01]. Decrease in GnRH and LH secretion during the pregnancy of rat may be controlled by constant expression of RFRP-3 mRNA and reduced expression of KiSS-1 mRNA in hypothalamus
Subject(s)
Animals, Laboratory , RNA, Messenger , Dorsomedial Hypothalamic Nucleus , Kisspeptins , Arcuate Nucleus of Hypothalamus , Rats, Sprague-Dawley , PregnancyABSTRACT
Herbal medicines are the major remedy in traditional medical systems and made a great contribution in maintaining human health and in preventing many infectious diseases. The present study was carried out to determine the potential antibacterial effect of ethanol extracts and essential oil of Marrubium vulgare L. against Staphylococcus aureus which is antibiotic resistant. All 17 strains of S. aureus isolated from nose and throat sample from 160 healthy subjects, hospital staffs and inpatient in the city of Zabol [Amir Al-Momenin hospital, Zabol, south-eastern Iran] were screened during years 2010-2011. In this study, the essential oil of Marrubium vulgare L. obtained by hydrodistillation was analyzed by gas chromatography coupled to mass spectrometry [GC-MS] in order to determine their chemical composition. The minimum inhibitory concentrations were investigated to characterize the antimicrobial activities of this essential oil and extract. Thirty-one components in the oil of Marrubium vulgare were identified. The results demonstrated that the major components of the essential oil were gamma-Eudesmol [11%], Germacrene [10%], D-Citronelly formate [10%], beta-Citronellol [8%], Geranyl tiglate [7.1%], Geranyl formate [6.02%]. The least MIC value of extract M. vulgare was 2.5 mg/mL and the highest MIC value of essential oil M. vulgare was 2.5 mg/mL. This investigation showed that the M. vulgare essential oil and extract has a potent antimicrobial activity against S. aureus. The present studies confirm the use of this essential oil and extract as antibacterial agent. Further research is required to evaluate the practical values of therapeutic applications
ABSTRACT
Tuberculosis is a global public health problem in the world. Microscopy of sputum smears is the most widely used method for diagnosing tuberculosis. However, many patients are smears negative for acid fast bacilli. Regarding the pathogenesis of the disease, the effectiveness of interferon-gamma [IFN-gamma] in bronchoalveolar fluid was investigated for the disease diagnosis. This descriptive study was performed at the Ali Ibn-e-Abitaleb hospital, Zahedan, between 2010 and 2012, to assess the role of IFN-gamma level in bronchoalveolar lavage in distinguishing tuberculosis from other pulmonary diseases. In patients who required fiberoptic bronchoscopy as indicated, bronchoalveolar lavage was analyzed in terms of smear acid-fast staining and cytology. The participants were divided into TB patients group [the BK smear of bronchoalveolar fluid or the culture was positive] and pulmonary non-TB patients group [the smear was negative]. Yet non-TB disease was definitively diagnosed by other means, as well. The fluid in each group was examined in terms of IFN-gamma. Then, Mean IFN-gamma levels in BALF were measured in these groups and then compared with each other. Eighty eight patients were enrolled in the study among which, 31 cases had TB and 57 patients suffered from pulmonary non-TB disease. Mean IFN-gamma was 2.85 +/- 4.17 pg/mL in pulmonary TB patients and 2.21 +/- 1.21 pg/mL in pulmonary non-TB patients. Lack of significant differences between the two groups in IFN-gamma indicate that this factor is not suitable for diagnosis of tuberculosis and differentiating it from other pulmonary diseases
ABSTRACT
Kisspeptin and RFamide-related peptide-3 [RFRP-3] are known to affect GnRH/luteinizing hormone [LH] in several species, including the rat. It has been hypothesized that GnRH/LH changes during the rat estrous cycle may result from changes in the expression of KiSS1 and RFRP-3 genes. Therefore, the present study investigates KiSS1 and RFRP-3 gene expression at the transcriptional level in the rat hypothalamus during the estrous cycle. In the present experimental study, 36 adult female Sprague-Dawley rats [3-4 months old] were used to study the expression of KiSS1 and RFRP-3 mRNA in the hypothalamus during the estrous cycle. Four rats were ovariectomized, whereas the remainder were allotted to four different phases of the estrous cycle [n=8 per estrus phase]. Rats were decapitated, and the hypothalami were immediately dissected and frozen in liquid nitrogen. Expressions of KiSS1 and RFRP-3 mRNAs were analyzed by real-time PCR. The expression of KiSS1 mRNA during estrus was lower than other phases of the cycle [p<0.01]. Expression of KiSS1 mRNA during the metestrus phase was lower than the proestrus phase [p<0.01]. The expression of RFRP-3 mRNA during proestrus was lower than the diestrus phase [p<0.01]. Results of the present study showed the role of coordinated expression of KiSS1 and RFRP-3 mRNA in the hypothalamus in the control of the rat estrous cycle