Transfection and cloning of genes for membrane antigens using the FACS

In order to facilitate cloning of genes for cell surface molecules, we cotransfected LTK^? mouse fibroblasts with thymidine kinase (TK) genes and total human or mouse DNA. TK⁺ cells, selected by growth in HAT medium, were stained with fluorochrome conjugated monoclonal antibodies or other fluorescent ligands which bind to one or another membrane differentiation antigen or receptor. We isolated fluorescent transfectants expressing these molecules using a fluorescence activated cell sorter (FACS). For some antigens, spontaneous gene amplification occurred. By repeated cycles of FACS sorting and regrowth we obtained high expressing clones. We then isolatedcDNA and genomic clones using selectedcDNA probes to screen phage withcDNA inserts. DNA from virtually any tissue source transfected equally well for the various molecules except for DNA from a trophoblast derived choriocarcinoma cell line which did not transfect for Leu-2.     

Fenfluramine treatment in infantile autism. Neurochemical, electrophysiological, and behavioral effects

As part of a multicenter, collaborative project, response to fenfluramine was assessed in 10 autistic outpatients. After 4 months of treatment, blood serotonin concentrations decreased an average of 60 per cent and returned to pretreatment levels after 2 months on placebo. This reduction was accompanied by a decrease in certain behavioral symptoms, including motor activity, distractibility, and mood disturbances. Baseline evoked potential recordings indicated that autistic patients tended to have a larger amplitude of the P3 component to frequent tones as compared to age-matched controls. A tendency toward "normalization" of the P3 effect was observed during the medication trial and during the final placebo period. Treatment response was not related to initial serotonin levels, and no major clinical side effects were associated with fenfluramine.     

Effects of priming duration on retention over the first 1 1/2 years of life

Exposing individuals to an isolated component (a prime) of a prior event alleviates its forgetting. Two experiments with 120 human infants between 3 and 18 months of age determined the minimum duration of a prime that can reactivate a forgotten memory and how long the reactivated memory persists. Infants learned an operant task, forgot it, were exposed to the prime, and later were tested for renewed retention. In Experiment 1, the minimum duration of an effective prime decreased logarithmically with age, but was always longer than the duration of a mere glance. In Experiment 2, the reactivated memory was forgotten twice as fast after a minimum-duration prime as after a full-length one, irrespective of priming delay and infant age. These data reveal that the minimum effective prime duration psychophysically equates the accessibility of forgotten memories. We conclude that priming is perceptually based with effects that are organized on a ratio (log) scale.     

Cytogenetic studies on the in-vitro genotoxicity of 4-nitroquinoline-1-oxide on human-lymphocytes

Using the UV-mimetic mutagen 4-nitroquinoline-1-oxide (4NQO) to induce genetic damage in human cells (lymphoblastoid lines and primary cultures of peripheral blood samples), chromosome aberrations were induced by treating the cells with 4NQO at 1 X 10(-5) M for 24 h. The overwhelming majority of chromosome aberrations was of the chromatid (S + G2) type instead of the chromosome (G1) type. The most common chromatid aberrations were simple breaks, isochromatid breaks, and chromatid exchanges. When the number of chromatid breaks per cell value was used as a measurement for 4NQO sensitivity, lymphoblastoid cells from a xeroderma pigmentosum patient showed the highest sensitivity, followed by the cells of two melanoma patients and normal persons. These preliminary results suggest that 4NQO may be employed to develop an assay system as a biomarker for determining UV sensitivity in the human population.     

The influence of abductor lever-arm changes after shoulder arthroplasty

Clinically, pain relief is usually satisfactory after shoulder arthroplasty, but the range of motion can be limited. The changes of the lever-arms of deltoid and supraspinatus muscles are considered to be important factors influencing postoperative active motion. After excluding patients with muscle weakness resulting from a neurologic condition, patients with chronic rotator-cuff tears, patients who did not follow rehabilitation guidelines, and patients with bilateral arthroplasties, 22 arthroplasty cases remained and were evaluated. Compared with the unaffected side, four radiologic parameters were assessed: (A) offset of humeral head, (B) lateralization of humeral head, (C) acromiohumeral interval, and (D) superoinferior migration of humeral head. The differences were measured and statistically analyzed. In summary: (1) The active and passive range of motion of abduction in scapular plane were 89.5° ± 26.1° and 109.7° ± 25.0°. (2) Parameters A, B, and C statistically changed (p < 0.05). (3) B value of the operated side was reduced below zero in six (27.3%). (4) There was a tendency to create medial shifting and inferior subluxation with the arthroplasty. This had a moderate adverse correlation to the active range of motion (correlation coefficient 0.498). (5) Before normalization for bone size, only C as a single parameter had moderate correlation with motion; after normalization, this decreased. (6) If combined parameters were assessed, A + C had a moderate correlation (correlation coefficient 0.435) with motion. (7) If an accurate anatomic relationship cannot be completely restored, A seemed more significant than any other parameters (p = 0.002); namely, shoulders with a smaller offset had a more limited active range of motion in abduction.     

A questionnaire to assess the generic and disease-specific health outcomes of patients with chronic hepatitis C

A 69-item questionnaire measuring generic functioning and well-being and disease-specific health outcomes was developed and tested using the pre-treatment data from patients with chronic hepatitis C (CHC) participating in two randomized trials of interferon -2b (n = 157). The questionnaire included all eight scales from the SF-36 and measures of nine other generic and disease-specific health concepts. Psychometric tests confirmed the assumptions underlying the construction and scoring of all generic and disease-specific scales. Cross-sectional tests of known groups validity showed that CHC patients scored worse on the generic scales than patients with other chronic conditions and worse than a healthy general population. The generic and disease-specific scale scores were lower in the presence of physical findings of CHC, as hypothesized, but only the physical functioning and bodily pain scales were linked to cirrhosis or extreme alanine aminotransferase (ALT) ratios. This instrument will be useful in studies of health outcome among patients with CHC, a condition whose health burden appears to have been underestimated in studies to date.     

Plasma selenium levels and risk of hepatocellular carcinoma among men with chronic hepatitis virus infection.

Both experimental and epidemiologic studies have linked a low dietary intake of selenium with an increased risk of cancer. The authors examined the association between plasma selenium levels and risk of hepatocellular carcinoma (HCC) among chronic carriers of hepatitis B and/or C virus in a cohort of 7,342 men in Taiwan who were recruited by personal interview and blood draw during 1988-1992. After these men were followed up for an average of 5.3 years, selenium levels in the stored plasma were measured by using hydride atomic absorption spectrometry for 69 incident HCC cases who were positive for hepatitis B surface antigen (HBsAg) and/or antibodies against hepatitis C virus (mostly HBsAg positive) and 139 matched, healthy controls who were HBsAg positive. Mean selenium levels were significantly lower in the HCC cases than in the HBsAg-positive controls (p = 0.01). Adjusted odds ratios of HCC for subjects in increasing quintiles of plasma selenium were 1.00, 0.52, 0.32, 0.19, and 0.62, respectively. The inverse association between plasma selenium levels and HCC was most striking among cigarette smokers and among subjects with low plasma levels of retinol or various carotenoids. There was no clear evidence for an interaction between selenium and alpha-tocopherol in relation to HCC risk.