Hybrid recombinant Omp 22, 25, and 31 immunodominant epitopes can be used for serodiagnosis of brucellosis.

Brucellosis is a well-known infectious disease in most parts of the world, especially in developing countries, common between humans and animals. Brucellosis is diagnosed by serological tests based on lipopolysaccharides (LPSs), which are bacterial cell wall antigens, and due to the similarities between LPSs antigens of some gram-negative bacterias, false-positive responses are inevitable. Alternatively, Outer membrane proteins (Omps), as antigenic conserved membrane proteins, can be used to diagnose brucellosis instead of LPS antigens. In this study, by using bioinformatics tools, linear B-cell epitopes were selected from Omp22, Omp25, and Omp31 antigens and fused with the rigid KP linker (K = Lysine, P=Proline). Designed gene cassette was cloned into pET-28a (+) vector and expressed recombinant protein was purified using Ni-NTA chromatography column and was confirmed with Poly-Histidine-HRP antibody. Finally, recombinant protein's seroreactivity with serum samples from 37 patients and 27 healthy individuals was evaluated by western blotting and enzyme-linked immunosorbent assay (ELISA) methods. Western blotting results showed high reactivity of the recombinant protein with serum samples of Brucella infected patients. ELISA results were analyzed using the receiver operating curve (ROC). Optical density cut-off point, accuracy, sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and Youden index J for recombinant protein were > 0.809, 84.37%,83.78%,88.89%,88.57%, 79.31% and 0.72 respectively. Western blotting and ELISA results showed that our recombinant protein has good sensitivity and specificity for the diagnosis of brucellosis.

Efficacy of doxycycline as add-on to interferon beta-1a in treatment of multiple sclerosis.

BACKGROUND: Available evidence shows that tetracycline family has cellular and molecular mechanisms to protect neurons and oligodendrocytes by modulating matrix metalloproteinases. We tried to compare the effectiveness of intramuscular and subcutaneous interferon beta-1a (INF-ß1a) in combination with oral doxycycline among patient with relapsing remitting multiple sclerosis (RRMS) and secondary progressive multiple sclerosis (SPMS). METHODS: A double-blind clinical trial study was conducted at Hamedan University of Medical and Health Sciences in Iran. Sixty patients with definite diagnosis of RRMS or SPMS were treated with doxycycline and 44 µg subcutaneous IFN-ß1a three times a week or 30 µg intramuscular IFN-ß1a once a week for six months. Neurologic examinations were performed monthly until the end of the treatment. Changes in expanded disability status scale (EDSS) scores, brain magnetic resonance images (MRIs), and frequency of receiving corticosteroid pulse were evaluated before and after the treatment. RESULTS: Women constituted 88.3% of the participants. The mean age of the patients was 32 years. The mean EDSS scores reduced from 4.5 to 3.0. Based on the frequency of receiving corticosteroid pulse, relapse rate decreased from 3.2 to 0.8. MRI showed that the number, volume, and activity of the lesions decreased among 13.3% of the participants, increased among 15%, and remained persistent among 71.7%. CONCLUSION: Combination of doxycycline and IFN-ß1a can decrease relapse rate and improve EDSS scores in patients with RRMS and SPMS. However, it does not affect MRI changes. Furthercontrolled clinical trials on greater number of patients with MS are needed to evaluate the efficacy of combination therapy.

Prevalence and location of petechial spots in well infants.

OBJECTIVE: To determine the prevalence and distribution of petechiae in well infants and to evaluate their relationship with age. DESIGN: 500 afebrile and well-appearing infants aged 1-12 months were examined for petechiae. The sites of the petechiae were noted on a skin map and the parent's explanation recorded. Data collection included the number of petechiae, age and birth rank. RESULTS: 10% of the infants had petechiae: 6.6% had one petechia and 3.4% more than one. Petechiae were on the trunk in 58% of infants, two of whom had them on more than one area. In 42% of the infants, the petechial spots were distributed on the head and neck as well as on the upper and lower extremities. Petechiae were significantly more common in infants over 6 months of age (68% vs 32%, p<0.05). During a 3-week follow-up period, all study infants remained well. CONCLUSION: 10% of infants less than 1 year of age were found to have petechiae. These data suggest that no further evaluation is necessary in cases where up to four isolated petechiae are observed, even if not confined to one body area, and the infant is well and afebrile.