[Cloning, sequencing and expression of HSP70 c-terminal domain in Mycobacterium avium subsp. paratuberculosis]

Heat shock proteins [HSPs] have been shown to act as an adjuvant when co-administered with different antigens, especially tumor antigens or antigens from infectious agents. C-terminal domain of Mycobacterium tuberculosis heat shock protein 70 [Hsp70], when fused to peptide antigens, provides a unique structure that is able to induce potent immune responses. In this study, aneukaryotic expression vector pEGFP-N1, containing C-terminal domain of Mycobacterium paratuberculosis HSP 70, Green Fluorescent Protein [GFP] gene in the plasmid construct, was designed for use as a reporter. With GFP system, expression of the target protein was evaluated in the cell culture. The nucleotide sequence of the cloned gene was revealed by sequencing. The protein expression of designed plasmid was also proved by reverse transcriptase polymerase chain reaction [RT-PCR]. Our eukaryotic expression vector [pEGFP-N1 -hsp70 c-terminal] was successfully constructed and HSP70 c-terminal domain protein was expressed effectively. The current experiment, as a basis for a new DNAvaccine design, can be used for the future studies on reverse vaccinology

Cloning and high level expression of bovine interferon gamma gene in eukaryotic cells [COS-7]

Interferon gamma [IFN-gamma] is one of the key cytokines in defining T helper 1 lymphocyte immune responses. In this study, the bovine IFN-gamma gene was cloned from spleen tissue RNA using the reverse transcription-polymerase chain reaction [RT-PCR]. IFN-gamma cDNA was sub-cloned and expressed in mammalian expression plasmid [pcDNA3.1[+]] under the control of the human cytomegalovirus [CMV] promoter. The predicted amino acid [aa] sequence of bovine IFN-gamma compared with corresponding known sequence from bovine [Bos taurus] was 100% identity and with ovine, caprine, camel, lama, equine, canine, feline, human, mice and chicken cytokine was 95, 95, 86, 83, 77, 75, 75, 61, 44 and 35%, respectively. Invitro expression of recombinant bovine IFN-gamma [rBoIFN-gamma] and secretion to culture medium was confirmed by ELISA test. Maximum expression of rBoIFN-gamma occurred at 96 and 144 h after transfection in COS-7 cells. These results showed that pcDNA3.1 expression vector and COS-7 cells transfected by diethylaminoethyl [DEAE]-dextran allowed the high level expression of bovine IFN-gamma gene and the release of protein in supernatant of cell culture

[Incidental sinus abnormalities in 256 patients referred for brain MRI]

Imaging abnormalities in the paranasal sinuses are regularly noted as incidental findings on MRI, however, little is known about their prevalence in the Iranian population. The purpose of this study was to classify these findings in the paranasal sinuses as seen on MRI and to investigate the prevalence, according to site and type of paranasal abnormality. In this cross-sectional study, the T2-weighted axial MRI of 256 patients with diseases unrelated to their paranasal sinuses were reviewed between May 2002 and June 2003. The findings were categorized according to the anatomic location and the imaging characteristics of the abnormality. The abnormalities recorded included total sinus opacification, mucoperiosteal thickening >/= 5mm, air fluid levels and retention cysts or polyps. Unilateral or bilateral involvement and septal deviation were also noted. A sinus was considered normal if it was fully aerated and no soft-tissue density was apparent within the cavity. Among our cases, 111 [43.5%] were male and 145 [56.5%] were female. Of these patients, abnormalities in one or more of the sinus groups were found in 110 subjects [42.9%], 55.5% of which were male and 44.5% were female [P=0.001]. Maxillary sinus abnormalities were observed in 66.4% of the patients, while ethmoid sinus abnormalities were found in 63.6%. Of the ethmoid abnormalities, 21% were found in the anterior section, 9% in the middle ethmoid, and 8% in the posterior ethmoid. The most common abnormality found was mucosal thickening. Among our cases, 23.4% had septal deviation, which was significantly higher among those with sinusitis [29% versus 19.1%; P<0.01]. Of those patients with sinus involvement, 16% were involved in the sphenoid sinus and 5% in the frontal sinus. The results obtained from the patients with sinus abnormality revealed that 85% suffered from cough, nasal obstruction, runny nose, facial pain and post nasal discharge and 24% had been diagnosed with chronic sinusitis by physician. Our results showed that there was a high rate of incidental abnormalities in the paranasal sinuses that are unrelated to the patient's presenting problems

[Effects of rapeseed meal and iodine supplement on broiler chicks]

To study the effects of rapeseed meal and iodine supplementation on performance, organs weight and some biochemical parameters of broiler chicks. Completely randomized design with 3 dietary treatments. One hundred and twenty one-day-old Arian broiler chicks [mixed male and female]. Chickens were categorized into 3 groups: soybean meal, rapeseed meal, and rapeseed meal plus iodine [2mg/kg]. feed intake, body weight gain and feed conversion ratio were determined at the end of starter, grower, finisher and whole priod states Moreover, the values of relative weight of liver, gallbladder, heart, spleen, gizzard, abdominal fat pad and carcass accompaning with serum concentrations of triiodothyronine, thyroxine, gamma glutamyltransferase and alanin aminotransferase were determined at 49 day of age. Significant differences among treatments determined by Duncan's multiple range tests. While rapeseed meal [RSM] reduced, body weight gain and feed conversion ratio, iodine supplement did not affect body weight gain and feed conversion ratio during starter, grower and whole period states. In contrast to iodine treatment the highest weight of gallbladder and its ratio to whole body weight were seen with RSM. The weight of abdominal fat pad and its ratio to whole body weight were reduced in RSM and RSM puls iodine supplementation. Iodine Supplementation had effect on the concentration of thyroxine, triiodothyronine gamm glutamyltransferase and alanin aminotransferase in RSM group. The complete replacement of rapeseed meal with soybean reduced performance and concentration of triiodothyronine hormone, and increased some organs weight, idoine supplementation had no effect on erfrmanc