Gypsy/Ty3-class retrotransposons integrated in the DNA of herring, tunicate, and echinoderms.

Eight new examples of retrotransposons of the Gypsy/Ty3 class have been identified in marine species. A 525-nt pol gene-coding region was amplified using degenerate primers from highly conserved regions and has extended the range of recognition of Gypsy/Ty3 far beyond those previously known. The following matrix shows the percentage AA divergence of the translations of this segment of the pol gene coding region. [table: see text] The underlines separate three groups of retrotransposons that can be recognized on the basis of this amino acid sequence. The new upper group shows surprising amino acid sequence similarity among members from the DNA of herring, sea urchin, starfish, and a tunicate. For example, the herring element differs by only 41% from the Ciona element and 46% from the sea urchin element. The group between the lines includes members close to previously known elements (marked by asterisks) and has so far been found only in sea urchins. The two upper groups differ from each other by 55-60% and yet members of both groups (e.g., Spr1 and Spr2) are integrated into the DNA of one species--S. purpuratus. Below the lower underline is listed the only known representative of a very distant group, which occurs in starfish DNA. In spite of large divergence, amino acid sequence comparisons indicate that all of the elements shown in the array are members of the LTR-containing class of retrotransposons that includes Gypsy of Drosophila and Ty3 of yeast. Of all known mobile elements this class shows the closest sequence similarity to retroviruses and has the same arrangement of genes as simpler retroviruses.

Rhinosporidiosis.

Rhinosporidiosis is the disease resulting from infection by Rhinosporidium seeberi, an organism endemic in the Indian sub-continent, but rarely seen in the United Kingdom. We describe the clinio-pathological features of a case presenting in a 14-year-old female.

Symptomatic cervical oesophageal ectopic (heterotopic) gastric mucosa treated with omeprazole.

A 60-year-old female patient with a 3-year history of nausea underwent oesophagogastroduodenoscopy. Red lesions were found in the cervical oesophagus: one of 2 cm and the other less than 1 cm. These lesions were biopsied and the larger lesion showed typical gastric mucosa. Following 40 mg/day omeprazole for 3 days all symptoms disappeared. Treatment was stopped after the patient remained symptom-free for 4 weeks.

Issues in cerebrospinal fluid management. Acid-fast bacillus smear and culture.

Meningeal tuberculosis is an uncommon disease in the United States with an annual incidence of fewer than 200 cases. This study evaluates three approaches to improving the use of the cerebrospinal (CSF) acid-fast bacillus (AFB) smear and culture procedure: (1) education alone; (2) optional screening by which physicians can select to have the AFB analysis stopped if the initial CSF findings are unremarkable; and (3) mandatory screening before the performance of all CSF AFB analyses. With education alone, the CSF AFB culture rate decreased from 20.6% of all CSF acquisitions to 15.7% (P less than 0.001); however, the effect may have been related to a decrease in all types of AFB testing. Optional screening had no impact on the AFB testing rate. Mandatory screening significantly decreased the CSF AFB rate to 6.7% (P less than 0.001), unrelated to changes in other types of AFB testing. Laboratories that employ mandatory screening should report the screening results immediately and have a mechanism whereby physicians can bypass the screen, providing CSF AFB analysis on unremarkable fluid from high-risk patients.

An adenoviral vector system for functional identification of nuclear receptor ligands.

A recombinant adenovirus system has been designed that confers glucocorticoid responsiveness upon infected cells in culture. Two mutually dependent viruses are required: a trans-activator virus containing the human glucocorticoid receptor transcription unit and a second receptor virus harboring a glucocorticoid response element linked to the firefly luciferase gene. Another reciprocal pair of viruses has been generated; one member expresses the rat thyroid hormone receptor alpha, while the other contains the luciferase gene regulated by a thyroid hormone-responsive DNA element. Corticosteroid- or thyroid hormone-induced transcription can be efficiently and accurately quantitated from cells coinfected with the appropriate complementary virus pair 20 h after infection in 96-well microtiter plates. This coinfection assay offers a convenient way to measure transcriptional activation by nuclear receptors and has certain key advantages over the commonly used cotransfection method. Its sensitivity and precision make it a practical approach to rapidly identify substances extracted from complex biological samples activating candidate "orphan" nuclear receptor molecules.

A system's architecture which dissociates management of shared data and end-user function.

An architecture for providing an institutional systems infrastructure is proposed. The architecture permits distributed applications while maintaining an integrated patient database.

Cerebrospinal fluid (CSF) TRAP. A method to improve CSF laboratory efficiency.

Establishment of a procedure termed cerebrospinal fluid (CSF) TRAP ("Transport and Rapid Accessioning for Additional Procedures") allows clinicians to appropriately store, at -75 degrees C, and rapidly access CSF specimens. The CSF TRAP enhances patient care by decreasing the need for repeat lumbar punctures and providing reserve fluid for the following: (1) further CSF testing; (2) repeating questionable test results; and (3) laboratory accidents. The CSF TRAP has been approved for third-party payment because it promotes efficient laboratory utilization by encouraging clinicians to review initial CSF findings before ordering low-yield CSF assays such as the venereal disease research laboratory (VDRL) and cryptococcal antigen latex agglutination tests. Currently, CSF TRAP samples are being obtained with 40% of all CSF acquisitions at the Duke University Medical Center. The availability of the CSF TRAP was associated with a significant decrease in the ordering of CSF VDRL and cryptococcal antigen assays (P less than 0.05); however, there was no significant change in the proportion of those studies being performed on normal CSF. The CSF TRAP procedure provides the framework for an overall restructuring of CSF testing that is being investigated.

Establishment of a CSF bank.

A bank of well-characterized CSF has been established by collecting and storing (-70 degrees C) CSF samples remaining after completion of routine clinical studies. Over 1,700 individual patient samples were collected during a 12-month period. A database derived largely from information down-loaded from existing hospital-based systems includes the results of individual CSF laboratory studies, in addition to the patient age, primary diagnoses, and details of any malignancy. CSF control material is used to verify storage conditions. The CSF bank supplies investigators with CSF handled in a standardized manner for more precise investigation of CNS disease.