A microwave technique for double indirect immunostaining of human brain tissue cultures with mouse monoclonal antibodies.

The use of 2 monoclonal antibodies during double immunohistochemistry would enable the use of a greater variety of antibody combinations. Here, we demonstrate a simple, cost effective method of double indirect immunostaining of cultured cells using primary antibodies from the same species. This method uses microwaving of cell samples immediately after the application of the first secondary antibody, and significantly reduces the level of nonspecific staining. This technique does not elute the antibodies, nor raise the sample temperature above 37 degrees C.     

Localization of the nonpalpable colonic lesion with intraoperative ultrasound

Localization of an nonpalpable colonic lesion at the time of colectomy usually requires intraoperative colonoscopy. The use of ultrasound to locate the lesion has not been described. A soft bowel clamp is placed above the expected location of the lesion and a catheter placed in the anus. Saline is then instilled into the colon and rectum. The lesion is located by ultrasound scan of the fluid filled colon with the probe placed on the serosal surface. Refinement of the technique was performed on resected colonic specimens. An in vivo trial was then performed with rapid and accurate localization of the lesion for resection. Intraoperative ultrasound of the colon can accurately localize nonpalpable colonic lesions and is an alternative to currently available techniques of localization. Received: 10 December 1997/Accepted: 11 March 1998     

AAV-mediated delivery of the caspase inhibitor XIAP protects against cisplatin ototoxicity.

HYPOTHESIS: Delivery of the gene encoding X-linked inhibitor of apoptosis (XIAP) using an adeno-associated viral (AAV) vector can protect against cisplatin-mediated ototoxicity. BACKGROUND: Cisplatin is a widely used chemotherapeutic agent with significant ototoxic side effects. One possible mechanism of toxicity is apoptotic death of many cochlear cell types. Acute treatment with inhibitors of caspases- enzymes critical for apoptosis- has been shown to prevent hearing loss in vivo, but is too short-acting for therapeutic use. Gene therapy provides a specific and chronic means of delivering potential therapeutic gents. Introducing an anti-apoptotic gene into the cochlea could provide long-term prophylaxis against the ototoxic effects of cisplatin. METHOD: Two groups of rats were treated with unilateral injection into the round window of AAV harboring a gene encoding either XIAP or green fluorescent protein (GFP). After at least two months of gene expression, auditory-brainstem-response (ABR) threshold shifts and outer-hair-cell (OHC) number were measured in these two groups of animals after 72-hour treatment with cisplatin. RESULTS: Consistent with previous reports, uninjected and AAV.GFP-injected ears displayed profound ABR threshold elevations and OHC loss after cisplatin treatment. Ears that had been injected with AAV encoding XIAP, however, were significantly protected from these effects: cisplatin-induced ABR-threshold shift and hair-cell loss were attenuated by as much as 78% and 45%, respectively, when compared with contralateral (untreated) ears. CONCLUSION: XIAP delivery to the cochlea can protect against the audiometric changes and hair-cell loss associated with cisplatin ototoxicity. The efficacy, specificity, and duration of the protective effects make this a potentially attractive therapeutic paradigm.     

VERIDICAL AND FALSE MEMORIES IN HEALTHY OLDER ADULTS AND IN DEMENTIA OF THE ALZHEIMER'S TYPE

Five groups of participants (young, healthy old, healthy old-old, very mild Dementia of the Alzheimer's Type, Mild Dementia of the Alzheimer's Type) studied and were tested on six 12-item lists of words selected from the DRM (Deese, 1959; Roediger & McDermott, 1995) materials. These lists of words strongly converged semantically on a nonpresented critical word. The results indicated that both veridical recall and recognition performance decreased both as a function of age of the participants and as a function of dementia severity. However, the recall and recognition of the highly related nonpresented items actually increased as a function of age, and only slightly decreased as a function of DAT. When false memory was considered as a proportion of veridical memory, there was a clear increase as a function of both age of the participants and as a function of disease severity. The results are discussed in terms of (a) age and DAT-related changes in attention and memory performance, and (b) the underlying mechanisms that produce false memories in the DRM paradigm.     

Interleukin-10 promoter polymorphisms in rheumatoid arthritis and Felty's syndrome

OBJECTIVE: To examine whether promoter polymorphisms associated with variation in interleukin-10 (IL-10) production are relevant to the development of rheumatoid arthritis (RA) or Felty's syndrome (FS). METHODS: DNA was obtained from 44 FS patients, 117 RA patients and 295 controls. The promoter region between -533 and - 1120 was amplified by polymerase chain reaction, and polymorphisms detected by restriction enzyme digest or sequence-specific oligonucleotide probing. RESULTS: We found no significant difference in allele or haplotype frequencies between the groups. CONCLUSION: There is no association between FS or RA and these recently identified IL-10 promoter polymorphisms. Other genetic or environmental factors could explain the alterations in IL-10 levels seen in these conditions.