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Biomedical subjects

C A Myers

Publications and source records attributed to C A Myers.

15 recordsLinked to original sources

Gene transfer in primary cultures of human hepatocytes.

Using liposomes as the mediator of DNA transfer, we were successful in the transfection of human hepatocytes isolated from surgical samples with an E. coli beta-galactosidase gene (beta-gal). A comparison of transfection efficiency showed that of the four promoters used, cytomegalovirus (CMV) promoter yielded higher transfection efficiencies than Rous sarcoma virus (RSV), Simian virus-40 (SV-40) and human alpha-1 antitrypsin (AAT) promoters. These studies represent the first report on the successful transfection of primary cultures of human hepatocytes.

Avian Sarcoma Viruses

A novel transcriptional enhancer is involved in the prolactin- and extracellular matrix-dependent regulation of beta-casein gene expression.

Lactogenic hormones and extracellular matrix (ECM) act synergistically to regulate beta-casein expression in culture. We have developed a functional subpopulation of the mouse mammary epithelial cell strain COMMA-1D (designated CID 9), which expresses high level of beta-casein, forms alveolar-like structures when plated onto the EHS tumor-derived matrix, and secretes beta-casein unidirectionally into a lumen. We have further shown that ECM- and prolactin-dependent regulations of beta-casein occur mainly at the transcriptional level and that 5' sequences play an important role in these regulations. To address the question of the nature of the DNA sequence requirements for such regulation, we analyzed the bovine beta-casein gene promoter in these cells. We now have located a 160-bp transcriptional enhancer (BCE1) within the 5' flanking region of the beta-casein gene. Using functional assays, we show that BCE1 contains responsive elements for prolactin- and ECM-dependent regulation. BCE1 placed upstream of a truncated and inactive beta-casein promoter (the shortest extending from -89 to +42 bp with regard to the transcription start site) reconstitutes a promoter even more potent than the intact promoter, which contains BCE1 in its normal context more than 1.5 kb upstream. This small fusion promoter also reconstitutes the normal pattern of regulation, including a requirement for both prolactin and ECM and a synergistic action of prolactin and hydrocortisone. By replacing the milk promoter with a heterologous viral promoter, we show that BCE1 participates in the prolactin- and ECM-mediated regulation.

Animals

Paradoxical effects of blood alcohol concentration charts.

BACKGROUND: This study was designed to determine the association between alcohol drinking and the possession of blood alcohol concentration charts. METHOD: A total of 30 college-age subjects participated in the study. Subjects were randomly assigned to either an experimental or a control group. All subjects answered a short entrance questionnaire to determine their height and weight. Those assigned to the experimental group were supplied a copy of a blood alcohol concentration chart and instructed in its use. The volume of alcoholic beverages consumed was surreptitiously counted for all subjects. Prior to leaving the premises the subjects completed an exit questionnaire which asked them to estimate the amount of alcohol they had consumed, whether they had driven to the pub, and whether they intended to drive away. The time spent in the pub was noted for each subject. RESULTS: Among those in the control group there was a tendency to overestimate the volume consumed, and for those in the experimental group, a tendency to more accurately estimate their consumption. Those given blood alcohol concentration charts consumed alcohol at a significantly higher rate than did those in the control group. CONCLUSION: A likely explanation for this outcome is that the chart served as a stimulus to prompt a drinker to more quickly achieve a blood alcohol level consistent with his/her drinking expectancies.

Adult

Extracellular matrix and hormones transcriptionally regulate bovine beta-casein 5' sequences in stably transfected mouse mammary cells.

Milk protein regulation involves synergistic action of lactogenic hormones and extracellular matrix (ECM). It is well established that substratum has a dramatic effect on morphology and function of mammary cells. The molecular mechanisms that regulate the ECM- and hormone-dependent gene expression, however, have not been resolved. To address this question, a subpopulation (designated CID 9) of the mouse mammary epithelial cell strain COMMA-1D has been developed in which more than 35% of the cells express beta-casein, form alveoli-like structures when plated onto a reconstituted basement membrane, and secrete beta-casein unidirectionally into a lumen. These cells were stably transfected with a series of chloramphenicol acetyltransferase (CAT) fusion genes to study transcriptional regulation of the bovine beta-casein gene. The expression of CAT in these lines demonstrated a striking matrix and hormone dependency (greater than 150-fold induction in some cases). This regulation occurred primarily at the transcriptional level and was dependent on the length of the 5' flanking region of the beta-casein promotor. Both matrix and hormonal control of transcription occurred within at least the first 1790 base pairs upstream and/or 42 base pairs downstream of the transcriptional initiation site. The ECM effect was independent of glucocorticoid stimulation. However, prolactin was essential and hydrocortisone further increased CAT expression. Endogenous beta-casein expression in these lines was similar to that of the parent CID 9 cells. Our data indicate the existence of matrix-dependent elements that regulate transcription.

Animals

Determination of peak trabecular bone density: interplay of dietary fiber, carbohydrate, and androgens.

To elucidate mechanisms linking nutrition and sex hormones to magnitude of peak trabecular bone density, we studied 11 normal women aged 19-21 y. Trabecular bone density was related inversely to dietary fiber (r = -0.69, p = 0.02) and carbohydrate (r = -0.70, p = 0.02) and directly to serum free-and-albumin-bound testosterone (fab T) (r = -0.70, p = 0.02) and total testosterone (total T) (r = 0.66, p = 0.03). Dietary fiber was correlated negatively with fab T (r = -0.74, p = 0.009), total T (r = -0.70, p = 0.02), and androstenedione (Adione) (r = -0.72, p = 0.01). Controlling for the effect of fab T or Adione weakened the relationship between dietary fiber and bone density and the relationship was no longer statistically significant. Conversely, controlling for sex hormones did not abolish the effect of carbohydrate on bone density. The contributions of fab T and carbohydrate to bone density were independent. These results suggest that dietary fiber may depress serum androgens which in turn decrease trabecular bone density. Carbohydrate may also depress bone density but independently of sex steroid hormones.

Adult

Effect of declining renal function on bone density in aging women.

The factors that are responsible for trabecular bone loss in aging women are not completely understood. To evaluate declining renal function as a possible factor, we studied 19 Caucasian women (average age 67) who were from 6 to 41 years postmenopausal. Trabecular bone density was quantitated by computerized tomography of the spine. Serum calcium, phosphorus, and creatinine were normal in all subjects. Creatinine clearance averaged 74 ml/min (range 38-122), decreased with age (r = -0.60, P = 0.003), and was inversely related to serum creatinine (r = -0.51, P = 0.01). Bivariate regression demonstrated that bone density decreased with age (r = -0.59, P = 0.004); controlling for the effect of creatinine clearance weakened this correlation to r = -0.45 (P = 0.03); controlling additionally for 1,25-dihydroxyvitamin D [1,25(OH)2D] and parathyroid hormone (PTH) reduced the correlation coefficient to r = -0.34 (P = 0.11). Bone density also decreased in direct proportion to the decrement in creatinine clearance (r = 0.44, P = 0.03); controlling for the effects of 1,25(OH)2D and PTH reduced this correlation coefficient to r = 0.34 (P = 0.11). These results suggest that occult renal insufficiency may contribute to bone loss in aging women, and that this effect may be mediated in part by 1,25(OH)2D and PTH. In this age group renal function should be assessed by measuring creatinine clearance rather than the serum creatinine concentration since renal insufficiency can be masked by apparently normal circulating creatinine levels.

Aged

Determinants of atraumatic vertebral fracture rates in menopausal women: biologic v mechanical factors.

In menopausal women, the susceptibility to atraumatic vertebral fractures is thought to be governed by both mechanical factors (as represented by bone density) and by biologic factors such as age, body size, and dietary calcium. Whether these biologic factors independently influence fracture rates beyond the effect of bone density is a matter of controversy. To compare the relative importance of mechanical and biologic factors on vertebral fractures, we elucidated the determinants of atraumatic compression fractures in 63 menopausal women who had no chronic diseases other than osteopenia. Trabecular bone density was determined by quantitative computerized tomography of the spine. Fracture frequency was expressed as the number of compressed vertebrae per person between T5 and L4. The analysis showed that fracture frequency, bone density, and the biologic factors were closely interrelated. Fracture frequency was inversely correlated to bone density [R2 (spline model) = .40, P less than .0001], body size (r = -.26, P = .05), and dietary calcium (r = -.28, P = .04), and directly correlated with age (r = .46, P = .0002). Bone density decreased with age (r = -.65, P less than .0001), increased with body size (r = .37, P = .004), and tended to increase with dietary calcium (r = .24, P = .08). After controlling for bone density, there was no perceptible residual relationship between fracture frequency and any of these biologic factors. We conclude that mechanical factors overshadow age, body size, and dietary calcium as determinants of vertebral fracture frequency. The apparent influence of these biologic factors on fracture frequency is explained by their surrogate effects on bone density.

Aged

In vitro evaluation of the cytotoxic potential of a novel man-made fiber, calcium sodium metaphosphate fiber (Phosphate Fiber).

As part of a comprehensive effort to evaluate the toxicological potential of calcium sodium metaphosphate fiber (Phosphate Fiber), the in vitro cytotoxicity of the fiber in cultured cells was studied. Two pulmonary-derived cell systems (rat alveolar macrophages, RAM; rat lung epithelial cells, LEC) and an established cell line (Chinese hamster ovary, CHO) were used. Release of lactate dehydrogenase (LDH) was used as an endpoint for cytotoxicity for all three cell types. In addition, inhibition of colony formation was used for CHO cells. The cytotoxicity of Phosphate Fiber was compared to a variety of mineral dusts and fibers including chrysotile asbestos, crocidolite asbestos, two glass fibers, calcium sulfate fiber, titanium dioxide, as well as the nonfibrous raw material, calcium sodium metaphosphate glass. Results with all three cell culture systems demonstrated that the Phosphate Fiber was less cytotoxic than the two asbestos fibers, similar in cytotoxicity to the glass fibers, and more cytotoxic than the calcium sulfate fiber and titanium dioxide. To further investigate the cytotoxicity of the Phosphate Fiber, it was fractionated by sedimentation into small and large fibers. The small Phosphate Fiber was found to be more cytotoxic and the large Phosphate Fiber to be less cytotoxic than the unfractionated Phosphate Fiber. The in vitro data suggest that Phosphate Fiber is less cytotoxic than asbestos, but further determination of safety can only be made after the in vivo data have been obtained.

Animals

A comparison of the risk of vertebral fracture in menopausal osteopenia and other metabolic disturbances.

The risk of atraumatic compression fracture in postmenopausal women increases as vertebral trabecular bone density decreases. To determine whether the risk is similar for patients who have other metabolic disorders, we compared eight-nine patients who had various disturbances affecting bone and sixty-three postmenopausal women who had no evidence of underlying disease. Trabecular bone density was measured by quantitative computed tomography of the lumbar spine. The relationship between frequency of fracture and bone density was modeled mathematically with spline threshold, quadratic polynomial, and decaying exponential functions. Analysis of covariance showed that the diagnostic category did not influence the relationship between frequency of fracture and bone density in any of the three models. We concluded that the risk of atraumatic compression fracture, as assessed by measurement of vertebral trabecular bone density using quantitative computerized tomography, is independent of the underlying metabolic disturbance.

Adult

Case report: unresponsive choroidal metastases.

1. The most common tumor of the choroid is metastatic tumor, with the lung as the primary site. Diagnosis may be difficult in distinguishing a choroidal melanoma from a metastatic lesion. 2. Careful metastatic evaluation may not identify an original primary tumor. Tests may not be sensitive enough to disclose adenocarcinoma until it has reached an advanced stage. 3. External beam radiation may not always be successful in treating choroidal metastases.

Adenocarcinoma, Mucinous

Choroidal osteoma.

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Choristoma