Differential role of the vagus in diurnal gene expression rhythms in the small intestine

The objective of this study was to examine the function of vagal innervation in maintaining diurnal rhythmicity in the expression of intestinal absorptive genes. Rats underwent truncal vagotomy and were maintained for 7 days on nighttime scheduled feeding (12-h light/12-h dark cycle). Vagotomized rats (V; n = 9) were pair-fed with sham-operated controls (S; n = 4). Unoperated normal rats (N; n = 6) were also included as controls. Half the rats were killed 3 h after lights on (ZT3; Zeitgeber Time, with lights-on considered ZT0) and the other half at ZT9, the time interval over which we have previously shown that sucrase and sugar transporter expression exhibits a significant anticipatory increase. RNA and protein extracted from mucosa of proximal jejunums were subjected to Northern and Western blot analyses to assess the increase in gene expression. Sham operation did not alter the normal diurnal rhythmicity of intestinal gene expression. Control rats (S plus N) exhibited the expected increase in RNA levels at ZT9 versus ZT3 for SGLT1 (4.5-fold), GLUT2 (5.3-fold), GLUT5 (4.1-fold), and sucrase (2.9-fold; P > 0.001 in all cases). In contrast, the induction in V rats was markedly blunted for GLUT2 (1.3-fold) and sucrase (1.5-fold) but not for SGLT1 (5.0-fold) or GLUT5 (4.2-fold). The mRNA levels for GLUT2 and sucrase at ZT9 were significantly lower in V rats versus controls (P < 0.001). GLUT2 and SGLT1 protein levels exhibited a parallel pattern: SGLT1 induction was 4.3-fold in control rats (P < 0.01) and 3.8-fold in V rats (P <0.01), whereas GLUT2 induction was 3.3-fold in control rats (P < 0.01) but only 1.4-fold in V rats (NS). Our results indicate that signaling through the vagus nerve is necessary to maintain the anticipatory induction pattern of GLUT2 and sucrase. The persistent rhythm in both SGLT1 and GLUT5 indicates that (1) diurnal induction of these genes is independent of vagal innervation and (2) the procedure did not cause an overall loss of intestinal function. Thus, entrainment of anticipatory diurnal gene expression in the intestine occurs via two separate pathways that are differentially dependent on vagal input.     

Astrocyte RNA in relation to neuronal RNA depletion in Alzheimer's disease

Summary A new double-staining procedure, in which the techniques of immunocytochemistry of glial fibrillary acidic protein (GFAP) and quantitative microdensitometry of azure B-RNA were combined, was used to study nucleic acid alterations in fibrous astrocytes in Alzheimer's disease (AD). RNA contents of GFAP-positive cells of the hippocampal endplate (Rose's H₃-H₅ fields) and the dentate gyrus molecular layer were determined in ten autopsy-proven AD patients (ages 51–88) and ten age-matched, non-demented control. In addition, RNA contents of pyramidal neurons of the endplate were examined. While there were no differences in RNA contents of astrocytes of either region between AD patients and controls, neuronal RNA was markedly depleted. These data suggest that astrocytes maintain protein synthetic capabilities in AD and that RNA loss is limited to the neuronal compartment.Supported by Grants 1P01-AG05119 and 1P50-AG05144 from the National Institutes of Health and by a Small Research Project Award from the University of Kentucky Medical Center     

Decreasing the level of translation initiation factor 4E with antisense rna causes reversal of ras-mediated transformation and tumorigenesis of cloned rat embryo fibroblasts

Transformation of cloned rat embryo fibroblasts (CREF) with the T24-ras oncogene results in loss of contact inhibition, growth in soft agar and tumor formation in nude mice. Previously we showed that in such cells (CREF T24), the phosphorylation rate of protein synthesis initiation factor 4E (elF-4E) is increased, correlating with an increase in the general rate of protein synthesis. In the present study, we have expressed antisense RNA complementary to elF-4E mRNA in CREF T24 cells using a stably integrated vector. Cells expressing antisense RNA (CREF T24/AS) contained 30-50% of the normal level of elF-4E and exhibited many of the properties of untransformed cells. CREF T24 had a spindle-shaped, refractile appearance, whereas CREF T24/AS grew in ordered, parallel patterns and exhibited contact inhibition similar to untransformed CREF. The rates of growth and protein synthesis in CREF T24/AS were decreased compared to CREF T24 but were not as low as in CREF. The efficiency of growth in soft agar was 11-fold lower for CREF T24/AS compared with CREF T24. The latency period for tumor formation in nude mice was increased from 8 days for CREF T24 to 17-27 days for CREF T24/AS and various clonal lines derived from them. Cell lines established from these CREF T24/AS-derived tumors were shown to have partially regained the elF-4E levels characteristic of CREF T24. These results demonstrate that many of the phenotypic alterations associated with ras-induced malignant transformation can be reversed by a moderate reduction of the translational initiation capacity and therefore may be mediated through a translational mechanism.     

Evaluation of the effectiveness of a community-based enriched model prenatal intervention project in the District of Columbia.

OBJECTIVE: To evaluate an enriched prenatal intervention program designed to reduce the risk of low birth weight. STUDY SETTING: Freestanding community-based prenatal intervention project located in a poor inner-city community, serving mostly African American women. STUDY DESIGN: All women less than 29 weeks pregnant were eligible to participate. They were compared to women who lived in neighborhoods with similar rates of poverty. DATA COLLECTION: The birth certificate was the source of data on maternal age, education, marital status, timing and frequency of prenatal care attendance, parity, gravidity, prior pregnancy terminations, fetal and child deaths, and birth weight. PRINCIPAL FINDINGS: Thirty-eight percent of the women who delivered live-born infants in the study area participated in the program. There were no differences in low- and very low birthweight rates in the study and comparison groups. In a secondary analysis comparing participants and nonparticipants in the study census tracts, participants were at higher risk for low and very low birth weight, and they adhered more closely to the schedule of prenatal visits than nonparticipants. Low- and very low birthweight rates were lower among participants than among nonparticipants and comparison women. CONCLUSION: The Better Babies Project did not have an effect on the overall low- and very low birthweight rates in the study census tracts. This was probably due to the low participation rates and the high population mobility.     

环孢菌素A阻断人HL－60抗药性细胞于G_1期而诱导敏感细胞凋亡

进一步研究了抗三尖杉酯碱的ＨＬ－６０细胞（ＨＲ２０）抗细胞凋亡的机制及该抗性和抗药性的关系。结果表明，环孢菌素Ａ（ＣｓＡ）２０，１０μｇ·ｍｌ￣（－１）诱导ＨＬ－６０细胞发生凋亡，而阻断ＨＲ２０细胞于Ｇ＿１期，就不能诱导细胞发生凋亡。低浓度的ＣｓＡ明显增加柔红霉素在ＨＲ２０细胞内的积聚，其逆转抗药性作用与阻断细胞周期运行无关。ＣｓＡ１０μｇ·ｍｌ￣（－１）处理ＨＲ２０细胞，可引起５０ｋＤａ的蛋白质高度磷酸化。结果提示：环孢菌素Ａ阻断抗三尖杉酯碱的ＨＬ－６０细胞于Ｇ＿１期，而诱导敏感的ＨＬ－６０细胞发生凋亡，其阻断作用与抗药性无关