Reactive astrocytes in neonate brain upregulate intermediate filament gene expression in response to axonal injury.

We have examined the injury response of astrocytes in the immature hamster brain in this study, focusing on alterations in the expression of glial fibrillary acidic protein (GFAP) and vimentin. In the adult CNS these two type III intermediate filament (IF) proteins have been shown to undergo robust increases in expression in response to axonal injury. Since injury to the immature CNS reportedly elicits less glial scar formation than adult brain injury, we examined the possibility that immature astrocytes respond differently than adult astrocytes to CNS injury with respect to IF gene expression. In situ hybridization using a 35S-labeled cDNA GFAP probe was done on brainstem sections obtained 2, 7 and 14 days after unilateral transection of the corticospinal tract in P8 hamster pups. The results indicated that substantial increases in GFAP mRNA were associated with the degenerating portion of the corticospinal tract by 2 days after axotomy and that the levels remained elevated for at least 14 days. Double-label immunofluorescence studies of this material suggested that GFAP as well as vimentin protein levels were also increased in many astrocytes in and around the degenerating corticospinal tract 2-14 days after axotomy. Most of the reactive astrocytes in the degenerating regions exhibited increases in GFAP and vimentin immunostaining but some vimentin-negative GFAP-positive reactive astrocytes were also observed, particularly in regions surrounding the actual degenerative zones.(ABSTRACT TRUNCATED AT 250 WORDS)

AUUUA-specific mRNA binding proteins in astrocytes.

Binding of ribonucleoproteins to specific regions of mRNA can alter mRNA stability. This level of posttranscriptional regulation has been shown to play a major role in gene expression of eukaryotic cells. This process involves the binding of ribonucleoproteins to specific region(s) of unstable, rapidly degrading mRNAs such as those found in various cytokines, lymphokines, and oncogenes, thereby increasing the mRNA's stability. In many instances the instability of the mRNA has been mapped to an AU-rich motif in the 3' untranslated region. We transcribed RNA molecules containing four reiterations of an AUUUA motif, and demonstrated with RNA- band shift experiments that the AUUUA motif complexes with phosphorylated AUUUA-specific 43-47 kDa mRNA binding protein(s) found in the cytosol of both rat brain and cultured rat astrocytes.

Phaeochromocytoma at the root of the mesentery.

A 65-year-old male reported with a freely mobile abdominal lump and hypertension. Though pre-operatively and intra-operatively, we have suspected the lump to be a mesenteric cyst, postoperative histopathological examination revealed it to be phaeochromocytoma which is rare. In view of its extra-adrenal site of presentation and rarity, this case is reported herein.

Puetz-Jeghers syndrome involving appendix.

Puetz-Jegher's syndrome is an autosomal dominant hereditary disease, which is characterized by hamartomatous polyposis and mucocutaneous pigmentation mainly over the circum-oral region. Patients with Peutz-Jeghers syndrome seek medical attention whenever there are complications such as intussusception, bleeding from the polyps, etc. Occasionally, gastrointestinal tract malignancies have been reported in Peutz-Jeghers syndrome. In this paper, we report a patient with Peutz-Jeghers syndrome who had multiple complications and polyposis involving the appendix, because involvement of the appendix is extremely rare in Peutz-Jeghers syndrome.

Gastric outlet obstruction due to duodenal tuberculosis: a case report.

A 17-year-old boy was operated on for gastric outlet obstruction; laparotomy revealed a mass with nodules in the first part of the duodenum and multiple mesenteric lymph nodes. The histopathological examination of the duodenal mass and the lymph nodes showed caseating tuberculosis. Because of the rarity of duodenal tuberculosis, the case is reported herewith.

Effects of ethanol on GLUT1 protein and gene expression in rat astrocytes.

Effects of ethanol on glucose transporter gene expression were examined in cultured rat astrocytes. Exposure to 50 or 100 mM ethanol for 18 hours significantly inhibited hexose uptake and reduced the number of glucose transporters, as indicated by binding studies with cytochalasin B. Indirect immunofluorescence and immunoperoxidase staining showed marked reduction of the GLUT1 glucose transporter by exposure to 100 mM ethanol for 5 or 18 hours, but no obvious change in response to 50 mM ethanol. Western blot analysis showed GLUT1 protein levels to be decreased by 52 +/- 12% (p < 0.05) after exposure to 100 mM ethanol for 18 hours. In situ hybridization histochemistry indicated an increase in steady-state GLUT1 mRNA in astrocytes exposed to 50 or 100 mM ethanol for 5 or 18 hours. Quantitation of GLUT1 mRNA levels by northern blot analysis showed that GLUT1 mRNA levels were increased by 59 and 112% in cells treated for 5 h with 50 and 100 mM ethanol, respectively. A similar effect was observed after treatment for 18 hours, but ethanol did not alter actin gene expression. Experiments using actinomycin D to block RNA synthesis suggest that this increase in steady-state mRNA level results from increased message stability. These results suggest that ethanol acts on GLUT1 gene expression at the post-transcriptional level.

Role of microtubules in glucose uptake by C6 glioma cells.

Drugs that influence tubulin function were used to investigate the role of microtubules in hexose uptake by C6 glioma cells. In C6 cells, colchicine and vinblastine (which inhibit tubulin polymerization) inhibited radioactive [3H]2-deoxy-D-glucose uptake by about 30%. Paclitaxel (which promotes tubulin polymerization) stimulated hexose uptake by about 25%. To further demonstrate that microtubules play a role in hexose uptake, C6 cells were transfected with GLUT1 cDNA and then challenged with 100 nM paclitaxel. In GLUT1-transfected cells paclitaxel stimulated 2-deoxy-D-glucose uptake by about 35%. To study the role of tubulin in agonist-stimulated hexose uptake, the effect of colchicine on carbachol-induced uptake was next examined. Hexose uptake was increased with carbachol in concentration-dependent manner which was abolished by pretreatment with colchicine. To examine the specificity of the inhibitory effect of colchicine on G protein-mediated signal transduction pathway, the influence of colchicine on insulin (which acts via tyrosine kinase pathway) stimulation of 2-deoxy-D-glucose uptake was investigated. Hexose uptake was increased by insulin in a concentration-dependent manner which was unaffected by pretreatment with colchicine. These results suggest that microtubules are involved in basal and carbachol-stimulated glucose uptake by C6 cells.

Glucose transporters and glucose utilization in rat brain after acute ethanol administration.

In the normal adult brain, glucose provides 90% of the energy requirements as well as substrate for nucleic acid and lipid synthesis. In the present study, effects of ethanol on glucose transporters (GLUT) and glucose utilization were examined in rat brain. Male Sprague-Dawley rats weighing 250-300 gms were given either ethanol 3 gm/kg BW or saline i.p. 4 hrs prior to the animal sacrifice and removal of the cerebral cortical tissue. The cortical plasma membranes analyzed by cytochalasin B binding assay showed a decrease in GLUT number but not in GLUT affinity in the ethanol treated rats as compared to the control rats. The estimated Ro values were 70 +/- 8.9 Vs 91 +/- 8.9 pmoles/mg protein (p < 0.05 N=4) and the estimated Kd values were 0.37 +/- 0.03 and 0.28 +/- 0.05 microM (p: NS) in ethanol and control experiments respectively. Immunoblots of purified cerebral plasma membranes and low density microsomal fraction showed 17% and 71% decrease for GLUTI and 54% and 21% (p<0.05 or less; n=6) for GLUT3 respectively in ethanol treated rats than in control animals. Immunofluoresence studies also showed reduction of GLUT1 immunoreactively in choroid plexus and cortical microvessels of ethanol treated rats as compared to control rats. The effect of ethanol on regional cerebral metabolic rates for glucose (CMR(Glc)) was studied using [6-(14)C] glucose and showed statistically insignificant decrease in brain glucose utilization. These data suggest that ethanol in-vivo decrease GLUT number and protein content in rat cerebral cortex.