Comparative Evaluation of Specific Antibody against Mycobacterium tuberculosis ESAT-6 Recombinant Antigen in Healthy Subject with Positive and Negative Skin Test.

Tuberculosis (TB) is an infectious disease caused by Mycobacterium tuberculosis (M. tuberculosis). The laboratory diagnosis of the disease includes various bacteriologic and immunologic methods. Despite the effectiveness of many of these methods in diagnosing active TB, their high cost and time-consuming nature have led researchers to adopt more accurate and rapid screening methods based on specific antigens for M. tuberculosis. The present study aimed to measure specific antibody serum levels against the early secretory antigenic target 6 kDa (ESAT-6) recombinant protein in healthy people and compare it to TB patients. The target population included 27 TB patients and 87 healthy individuals with no clinical TB symptoms. The healthy population was divided into two groups, including positive purified protein derivative (PPD) and negative PPD (35 and 52 people, respectively), using the Tuberculin skin test. The specific antibody level against the ESAT-6 recombinant antigen and the PPD protein was measured using an indirect Enzyme-Linked Immunosorbent Assay (ELISA) test. The results of the study showed that the majority of the healthy population with no symptoms of clinical TB and having negative skin test results did not have antibodies against the recombinant ESAT-6 (98%) and PPD (96%) antigens. On the other hand, there was a high level of the specific antibody of the ESAT-6 recombinant and PPD antigens in TB patients (77%). It is notable that in people with positive skin test results, the level of the antibody against the ESAT-6 recombinant antigen and PPD antigen was 94%. The results demonstrated that the ELISA method based on the measurement of antibodies against the ESAT-6 recombinant antigen can be a proper diagnostic method for rapid and accurate screening of healthy from infected people.

Effects of Adjuvant and Immunization Route on Antibody Responses against Naja Naja oxiana Venom.

Naja naja oxiana (NNO) is one of the important venomous species in Iran. The current snakebite treatment is antivenom therapy that deals with hyper immunization of horses with crude or fractionated snake venom plus traditional adjuvants, like Freund's adjuvant. For improvement of antivenom production, it has been suggested to use different adjuvant systems or immunization procedures. In this study, humoral immune responses against immunogenic fractions of NNO venom (NNO3 and NNO4) and crude venom have been compared by usage of different adjuvant and immunization routes. Additionally, a new indirect enzyme-linked immunosorbent assay (ELISA) was set up for the detection of specific antivenom antibodies. This study was conducted on six different groups of female Dutch rabbits that were hyperimmunized using crude and fractionated NNO venom, along with Freund's and MF59 adjuvants through subcutaneous or intramuscular route. The immunization was performed four times with 10-day intervals and the levels of specific antibodiees were detected by indirect ELISA. The statistical analysis reveals a negligible variation in the antivenom titers among the venom-inoculated groups, regardless of the adjuvant type or the immunization route. Finally, it was concluded that the fractions are efficient for antivenom production, and it is possible to use MF59 adjuvant via subcutaneous routes as an alternative to Freund's adjuvants considering its fair immunopotentiation capacity and safety in animals.

Novel Applications of Immuno-bioinformatics in Vaccine and Bio-product Developments at Research Institutes.

There are many challenges in the field of public health sciences. Rational decisions are required in order to treat different diseases, gain knowledge and wealth regarding research, and produce biological or synthetic products. Various advances in the basic laboratory science, computer science, and the engineering of biological production processes can help solve the occurring problems. Bioinformatics is defined as a field of science combined of biology, mathematics, physics, chemistry, and computer sciences. Recently, bioinformatics has been extensively used in the designing of the epitope, vaccines, antibodies, adjuvants, diagnostic kits, and therapeutic purposes (e.g., proteins, peptides, or small molecules). Moreover, bioinformatics includes chemoinformatics that has been employed to produce various biological or chemical products to target and combat pathogens. Bioinformatics is involved in other areas of data analysis and prediction, such as structural biology, system biology, phylogeny, population genetics, and next-generation data sequencing. To the best of our knowledge, no published study coherently described the benefits of bioinformatics fields applied for medication development or diagnostic aims in bio-productive and pharmaceutical/vaccine companies. Therefore, in the current review, we attempted to present the available bioinformatics resources, practical experiences, and other findings in the mentioned field along with providing a harmonized and applied model(s). The key points presented in the current review may help to elevate production and reduce the costs for the development of novel vaccines, medicines, and antibodies. In addition, these methods can facilitate the identification of organisms and may guarantee the quality of biological products.

Production of MPT-64 recombinant protein from virulent strain of Mycobacterium bovis.

Tuberculosis (TB) is a zoonotic infectious disease common to humans and animals which has been caused by a rod shaped, acid fast bacterium, called Mycobacterium bovis. The rapid and sensitive detection is a great challenge for TB diagnosis. The virulent strains of Mycobacterium tuberculosis complex (MTBC) have 16 different regions of difference (RD) in their genome which encode some important antigens. The major protein of M. bovis 64 (MPT-64) is one of the main immune-stimulating antigens which are encode by RD-2 region. The aim of the present study was cloning, expression and purification of MPT-64 as a protein antigen of M. bovis in a prokaryotic system for the usage in the future diagnostic studies. In this experimental study, the mpt-64 gene with 687 bp has been proliferated from M. bovis whole genome by polymerase chain reaction (PCR) method. The PCR product has been digested by BamHI and HindIII restriction enzymes and cloned into pQE-30 plasmid. The recombinant protein has been expressed in the Escherichia coli M15 with induction by isopropyl-ß-D-thiogalactopyranoside (IPTG). The expressed protein was analyzed on SDS-PAGE, and purified with Nitrilotriacetic acid (Ni-NTA) column. Finally, its biological properties were confirmed in Western blotting method using specific antibodies. Data showed the successful cloning of mpt-64 gene (as a 687 bp segment) in expression vector. The MPT-64 recombinant protein was ideally expressed and purified as a 24 kDa protein. The result of this study indicated that MPT-64 recombinant protein (24 kDa) has been successfully expressed and purified in a prokaryotic system, so this protein could be used for differential diagnosis of pathogenic and non-pathogenic Mycobacterium, in suspected BTB cases.

The effect of feeding excess arginine on lipogenic gene expression and growth performance in broilers.

1. Reducing excess fat accretion is important for both human health and animal production. The present study was conducted to investigate the effects of arginine (Arg) on the regulation of lipogenic gene expression and on growth performance. 2. One-d-old female broiler chicks (Ross, n = 192) were used in a completely randomised design with 4 dietary treatments in which diets included 100% (CTL), 153% (LArg), 168% (MArg) and 183% (HArg) of the recommended concentration of digestible Arg. 3. Results showed that high concentrations of Arg improved body weight gain, feed efficiency, meat production, fat and crude protein content of breast muscle and plasma thyroid hormones. Conversely, abdominal fat, cholesterol, triglyceride and urea were lower with higher concentrations of Arg. Dietary arginine increased lipogenic gene expression in muscles, while decreasing those in adipose tissue and liver. 4. It was concluded that increasing Arg in the diet reduced abdominal fat content, enhanced intramuscular fat and increased muscle and protein gain. Furthermore, Arg supplementation at the MArg concentration improved growth performance, and at HArg had the greatest effect on fat reduction.