Use of the recombinant 38-kDa antigen of Mycobacterium tuberculosis as an immunogen for specific antisera production

The Mycobacterium tuberculosis 38-kDa protein antigen is one of the secreted immunodominant antigens showing high immunogenicity at B-cell and T-cell levels. Although monoclonal antibodies to this antigen have been produced, specific polyclonal antisera is required for standardization of specific immunodiagnostic assays. This protein has been overexpressed and purified from recombinant Escherichia coli using an inducible vector system. During each stage of expression and purification, the recombinant protein was used to immunize mice and rabbits by several methods: 1) as overexpressed protein present as inclusion bodies in recombinant E. coli; 2) embedded in a polyacrylamide gel; 3) fixed to a solid-phase nitrocellulose membrane and 4) emulsified with an adjuvant. All strategies yielded specific antisera as determined by enzyme-linked immunosorbent assay (ELISA) and immunoblot analyses. The results obtained, both quantitative (ELISA) and qualitative (immunoblot) demonstrate that the purified recombinant antigen retains its antigenicity and immunogenicity throughout the various steps in the process of expression and purification and serves as a potent antigen for production of specific antisera to be used in immunoassays.     

Effects of pancreatic polypeptide amylin on ulceration and acid gastric secretion

The effects of the pancreatic polypeptide amyline on ulceration and acid gastric secretion were studied in rat experiments. Pyloric ligation was used as a model of ulceration. Amyline administration caused significantly less gastric mucosal damage in response to pyloric ligation. The severity of gastric mucosal damage averaged 47 +/- 13 mm2 in the control group and 25 +/- 11 mm2 (p < 0.005). The rate of acid gastric secretion in the animals whose pylorus had been ligated as judged by the pH of gastric content was significantly higher than that in the controls (2.87 +/- 0.22 and 2.34 +/- 0.17 (p = 0.05). It is concluded that amyline has a noticeable effect on the gastric mucosa. It is suggested that suppressed acid gastric secretion, i.e. reduced influence of aggressive agents on the gastric mucosa, is a mechanism of antiulcerative action of the peptide.     

Clonal relationships among high-level penicillin-resistant Streptococcus pneumoniae in the United States

Emu antibody responses to avian influenza virus (AIV) infection were evaluated by the competitive enzyme-linked immunosorbent assay (C-ELISA), agar gel immunodiffusion (AGID) and hemagglutination inhibition (HI) tests. All birds infected with AIV H5N1, H5N3, or H7N7 developed antinucleoprotein (NP) antibodies as early as 7 days postinfection as detected by the C-ELISA. The responses lasted 49 days for the emus receiving H5N3 and at least 56 days for emus receiving the other two viruses. By evaluating 50 emu field serum samples, the C-ELISA was found more sensitive than the AGID test for the detection of anti-NP antibodies. This study indicates that emus experimentally infected with AIV developed antibody responses that can be detected by C-ELISA, AGID, and HI tests. The results from this and our previous studies demonstrate the use of the C-ELISA as a substitute for the AGID test in a routine serodiagnostic screening for detection of antibodies to AIV infection in multiple avian species.     

Reconstructive and plastic microsurgery in the treatment of combat trauma to the extremities

In the department of microsurgery of A.A. Vishnevsky Central Hospital the transplantation and transposition of the at the gunshot injuries of the extremities have begun to execute since 1990. For this time 94 surgical interventions have been conducted at 88 wounded: 57 transplantations and 37 transpositions of the bloody tissues. The transplantation and transposition of the vascularized complexes of tissues with application of microsurgical technique is the effective method of the treatment of the wounded in the extremities. In the conditions of the local armed conflicts it is necessary to provide deployment of the department of microsurgery at stages of the specialized medical care.     

A comparative study of passive haemagglutination methods for the detection of hepatitis B surface antigen in routine hospital practice

Two passive haemagglutination methods for detecting HBsAg were compared. In general, the method using turkey erythrocytes was found preferable to the method employing sheep cells since it is more rapid and more sensitive, and less frequently gave rise to false positive reactions with sera from staff, blood donors, and patients not receiving haemodialysis. The turkey cell test gives rise to more false positive screen tests than the sheep cell test when monitoring renal dialysis patients since approximately 10% of the sera of these patients were found to contain turkey cell agglutinins, but this presents no particular difficulty if the recommended absorption procedures are used.     

Baseball implant. A method of secondary insertion of an intraorbital implant

A surgical technique for secondary emplacement of an orbital implant is described in which a spherical implant encased in a scleral homograft is placed in the orbit through a transconjuctival incision and sutured to the superior part of the periosteum.     

Parathyroid hormone (PTH) stimulates adrenocorticotropin release in AtT-20 cells stably expressing a common receptor for PTH and PTH-related peptide

Complementary DNA encoding a rat bone PTH/PTHrP receptor was stably expressed in the murine corticotroph cell line, AtT-20. Several clones, expressing variable numbers of PTH/PTHrP receptors, were developed. In contrast to the relatively low binding affinity (apparent Kd = 15 nM) observed in COS-7 cells transiently expressing the PTH/PTHrP receptor, all AtT-20 stable transfectants bound [Nle8,18,Tyr34]bPTH(1-34)NH2 (NlePTH) with an affinity that was indistinguishable from that observed in ROS 17/2.8 cells expressing native PTH/PTHrP receptors. Additionally, NlePTH dramatically increased cAMP accumulation and ACTH release in AtT-20 cells expressing the PTH/PTHrP receptor with an ED50 of 0.6 +/- 0.3 and 0.3 +/- 0.1 nM, respectively. The high binding affinity and the high efficacy of NlePTH in stimulating cAMP accumulation and ACTH release indicate that the PTH/PTHrP receptor is efficiently coupled to the intracellular signalling system responsible for stimulation of ACTH release in AtT-20 cells. No additivity of cAMP accumulation or of ACTH release was observed when these cells were treated with maximally active concentrations of both NlePTH and CRF. This suggests that the receptors for both of these hormones share the same intracellular effectors, and that intracellular signaling in AtT-20 cells is not compartmentalized. Additionally, the ability of NlePTH to stimulate ACTH release in AtT-20 cells, a function that is normally performed by CRF, demonstrates promiscuity between activated receptors and distal biological functions.