Characterization of the CAMPATH-1 (CDw52) antigen: biochemical analysis and cDNA cloning reveal an unusually small peptide backbone.

The CAMPATH-1 (CDw52) antigen has been purified from human spleen. The antigenic epitope is heat stable but sensitive to mild alkali treatment. Experiments with phosphatidylinositol-specific phospholipase C indicate that it is anchored by a glycosylphosphatidylinositol (GPI) anchor. An N-terminal sequence of 11 amino acids was determined, followed by an abrupt stop. Using short overlapping mixed oligonucleotide primers, cDNA synthesized from the mRNA of a human B cell line was amplified by the polymerase chain reaction. The product was used to isolate cDNA clones and the full amino acid sequence of the CAMPATH-1 antigen was deduced. It consists of 37 amino acid residues plus a 24-residue signal peptide. It has all the features expected for a GPI-anchored membrane protein except that the predicted mature protein is remarkably short, comprising no more than 18 residues and possibly as few as 12 (depending on the GPI linkage site). Potential attachment sites for carbohydrate are present and it is shown that the antigen contains N-linked oligosaccharide(s). This structure accounts for the known properties of the antigen, though the exact reasons why it is such a good target for cell lysis in vitro and in vivo are not yet clear.     

Suppression of atherogenesis by n-3 fatty acids in the cholesterol-fed rabbit.

The effects of n-3 fatty-acid supplementation on serum lipids, platelet aggregation, and the development of atherosclerotic lesions were studied in the cholesterol-fed rabbit. Serum total cholesterol and LDL cholesterol values were significantly reduced in comparison with those of the nonsupplemented cholesterol-fed group (p less than 0.005, p less than 0.0025, respectively), though still higher than those of the control group (p less than 0.0025, p less than 0.0125 respectively). Platelet aggregation was reduced below that of the cholesterol-fed and the control levels (p less than 0.0005, p less than 0.0025, respectively). The endothelial injury encountered in cholesterol-fed rabbits was inhibited in the supplemented group. It is concluded that n-3 fatty acids suppress atherogenesis in this animal model by interfering with platelet aggregation and lipid metabolism.     

Campath-1M--prophylactic use after kidney transplantation. A randomized controlled clinical trial

Campath-1M is a rat monoclonal IgM antibody that binds human complement and recognizes virtually all peripheral human mononuclear cells. It is known to be effective in T cell depletion of bone marrow grafts, and encouraging results were obtained in a pilot study in which the antibody was used in prevention and treatment of rejection of kidney, pancreas, and liver allografts. In this randomized controlled clinical trial, Campath-1M has been evaluated as a prophylactic agent following renal allografting. It is shown that patients who received a 10-day course of the antibody immediately postoperatively, in addition to standard therapy with high-dose cyclosporine (17 mg/kg), experienced a significantly lower incidence of early acute cellular rejection than control patients who received cyclosporine alone. There was no evidence of "rebound" rejection following the end of antibody treatment to suggest that rejection had merely been delayed. However, patients who received this additional immunosuppression experienced a significantly higher incidence of serious infections than controls, this negating any benefit from the treatment in terms of graft survival. Thus, a monoclonal antibody of broad specificity directed against lymphocytes may be effective as a prophylactic agent after organ transplantation but its use should be accompanied by a reduction in other immunosuppressive drugs.     

Cathepsin-D expression in cervical carcinoma and its prognostic significance

An immunohistochemical study was made of cathepsin-D protein expression in each of the three main types of uterine cervical carcinoma (squamous carcinoma, adenosquamous carcinoma and adenocarcinoma) with particular reference to lymph node status and prognosis. Of the 61 cases, 54.1% showed cytoplasmic staining in more than 2.5% of tumour cells counted. Cathepsin-D expression was significantly higher in adenocarcinoma (mean -3.128) than in squamous carcinoma and adenosquamous carcinoma (mean –3.709,P=0.047 using logit transformation). Cathepsin-D had no prognostic value in any of the three tumour types. No relationship was found between cathepsin-D staining and lymph node status and there was no advantage in adding cathepsin-D values to lymph node status. These results suggest that immunostaining for cathepsin-D protein expression is unlikely to be of use as a prognostic marker.     

Rapid identification by the Micro-ID system of Enterobacteriaceae detected by urine screening.

Previous studies have demonstrated that organisms detected by urine screening can be processed for rapid identification and antimicrobial susceptibility testing directly from urine or urine screening broth. In the present study, an improved method for processing such specimens was evaluated. Organisms were harvested by centrifugation from positive urine screening broth, and inocula were prepared for rapid identification by the Micro-ID system and rapid susceptibility testing by the Autobac system. Nearly 2,500 urine specimens were analyzed by urine screening, and 206 specimens had significant growth of gram-negative, oxidase-negative bacilli. These organisms, prepared by the centrifugation procedure, were identified and tested for susceptibility to antimicrobial agents. For comparison, identifications by the Micro-ID system and antimicrobial susceptibility tests by the Autobac system were performed on the same organisms the next day with inocula prepared from colonies growing from standard urine cultures. The results demonstrated that 95% of the organisms were correctly identified by this procedure, and susceptibility testing by the rapid method gave results in 94% agreement with the standard method. These results demonstrate that organisms detected by urine screening can be accurately identified and tested for antimicrobial susceptibility after centrifugation from urine screening broth. This system provides a practical procedure or same-day reporting of urine culture results.     

Rapid Semiautomated Screening and Processing of Urine Specimens

A rapid urine culture procedure was evaluated in which positive urines were detected by using light-scatter photometry (Autobac). Specimens were analyzed at 3, 5, and 6 h. Specimens detected as positive at 3 h were then further evaluated by a direct 3-h susceptibility procedure (Autobac) and by a 4-h identification procedure (Micro-ID). Of 949 specimens, 175 had >10⁵ colony-forming units per ml by colony count. Of these latter specimens, 75.4% had been detected by 3 h, and 95.4% were detected by 6 h. Of specimens positive by Autobac at 3 h, 96% (95.7%) had >10⁵ colony-forming units per ml. If pure by Gram stain, those positive specimens were inoculated to direct susceptibility and identification systems. When direct Autobac susceptibilities were compared with the standard Autobac method done from the plate the following day, discrepancy rates were 1.3% very major, 2.1% major, and 7.4% total. The direct identifications were 94% (94.2%) correct when using the Micro-ID manual and a collection of octal patterns unique to this system, in which urine/broth culture inoculum was employed instead of the usual organism colony suspension. Those urine specimens negative after screening at 3 h were evaluated at 5 and 6 h, and an additional 126 specimens were detected as positive. These were then processed by routine plate inoculation, due to the limitations of the work day. By 6 h, 95.4% of specimens with >10⁵ colony-forming units per ml were detected. The 4.6% false-negative results consisted of patients on antibiotics, or slowly growing bacteria suspected of being distal urethral contaminants. Thus, 83.5% of the urine cultures received by 9:00 a.m. (10.6% 3-h positives and 72.9% negative at 6 h) could be evaluated and reported within one 8-h work day.     

Isolation of low-frequency class-switch variants from rat hybrid myelomas

Class-switch variants have been isolated from rat-rat hybrid myelomas by sib selection using a simple assay based on red cell-labelled antiglobulins. The variants detected are consistent with the gene order deduced from molecular cloning. They appear to arise spontaneously at a rate approximately ten-fold lower than for mouse cell lines but the rate of switching back to the parental isotype is substantial in comparison. An IgG2b variant antibody having the same specificity as CAMPATH-1 for human lymphocytes and monocytes is active in antibody-dependent cell-mediated killing (unlike the parental IgG2a) and may prove to be a valuable therapeutic antibody for immunosuppression and treatment of leukaemia and lymphoma.     

Infective agents in fixed human cadavers: a brief review and suggested guidelines

Cadavers remain a principal teaching tool for anatomists and medical educators teaching gross anatomy. Infectious pathogens in cadavers that present particular risks include Mycobacterium tuberculosis, hepatitis B and C, the AIDS virus HIV, and prions that cause transmissible spongiform encephalopathies such as Creutzfeldt-Jakob disease (CJD) and Gerstmann-Straussler-Scheinker syndrome (GSS). It is often claimed that fixatives are effective in inactivation of these agents. Unfortunately cadavers, even though they are fixed, may still pose infection hazards to those who handle them. Specific safety precautions are necessary to avoid accidental disease transmission from cadavers before and during dissection and to decontaminate the local environment afterward. In this brief review, we describe the infectious pathogens that can be detected in cadavers and suggest safety guidelines for the protection of all who handle cadavers against infectious hazards.