Maturation of tumor vasculature by interferon-beta disrupts the vascular niche of glioma stem cells.

BACKGROUND: The vascular niche necessary for cancer stem cell maintenance is a potential target for cancer therapy. MATERIALS AND METHODS: Human glioma xenografts were treated with IFN-ß delivered systemically via a liver-targeted, adeno-associated viral vector. The vascular niche was examined with immunofluorescence for glioma stem cells, endothelial cells, and perivascular cells. RESULTS: Although IFN-ß was not directly toxic to glioma stem cells in vitro, IFN-ß decreased tumor size and the number of stem cells recovered in both heterotopic and orthotopic models. Treatment with IFN-ß increased perivascular cells investing the tumor vasculature (6-fold) distancing stem cells from endothelial cells. Additionally, vascular smooth muscle cells co-cultured between stem cells and endothelial cells decreased stem cell recovery. CONCLUSION: Continuous delivery of IFN-ß decreased the number of stem cells in glioma xenografts by disrupting the vascular niche through an increase in perivascular cells, which created a barrier between the glioma stem cells and the endothelial cells.

Multimedia quality of life assessment: advances with FLAIR.

Assessing impact of functional dependency on quality of life (QOL) among older adults can provide an in-depth understanding of health preferences. Utilities as a measure of preferences are necessary in conducting cost-effectiveness evaluations of healthcare interventions designed to improve overall QOL. We describe further development of a multimedia utility elicitation instrument that is highly portable and easily accessible. An earlier version, FLAIR1, introduced features designed for older adult, computer inexperienced users. FLAIR2 includes modifications such as migration to a web-based platform, consistency checks, audio/visual updates, and more response methods. As compared with FLAIR1, more FLAIR2 respondents (n=318) preferred using the computer and found the computer program to be enjoyable, easy to use, and easily understood. There were also fewer inconsistencies among FLAIR2 respondents. FLAIR2 enhancements have increased portability, minimized invariance and inconsistency, and produced a more user friendly design.

Intraoperative ultrasound and prophylactic ursodiol for gallstone prevention following laparoscopic gastric bypass.

BACKGROUND: Previous studies have shown that ursodiol decreases gallstone formation from 32% to 2% following open gastric bypass, but no data exist on laparoscopic Roux-en-Y gastric bypass (LRYGB) using intraoperative ultrasound (IOUS) screening. METHODS: LRYGB with IOUS were performed on 195 consecutive patients. Patients with gallstones underwent simultaneous cholecystectomy, and patients without gallstones were prescribed ursodiol, 300 mg twice daily, for 6 month. Follow-up survey and ultrasound. RESULTS: Of 195 patients, 44 (23%) had had a prior cholecystectomy, 21 (11%) underwent a simultaneous cholecystectomy, 129 (66%) had gallbladders left intact, and one (0.5%) false negative IOUS was excluded. Of 69 patients with ultrasound and survey follow-up (mean, 10 months), 19 (28%) developed gallstones seven with symptoms), and 50 (72%) were gallstone free. Forty-one percent of patients were compliant with ursodiol. There was no difference in compliance between patients with and without gallstones. In patients with gallstones, all of the symptomatic patients were noncompliant, whereas none of the compliant patients developed symptoms. Medication side-effects occurred in 17 of 69 patients (25%). CONCLUSIONS: IOUS during LRYGB efficiently screens for gallstones, and selective cholecystectomy followed by prophylactic ursodiol results in low morbidity. Improvements in compliance may lower the incidence of postoperative gallstone formation.

Clinical predictors of leak after laparoscopic Roux-en-Y gastric bypass for morbid obesity.

BACKGROUND: Gastrointestinal leak is a complication of laparoscopic Roux-en-Y gastric bypass (LRYGB). Contrast studies may underdiagnose leaks, forcing surgeons to rely solely on clinical data. This study was designed to evaluate various clinical signs for detecting leakage after LRYGB. METHODS: We retrospectively reviewed 210 consecutive patients who underwent LRYGB between April 1999 and September 2001. There were nine documented leaks (4.3%). Clinical signs between patients with leaks (group 1) and those without leaks (group 2) were compared using univariate and multivariate logistic regression analysis. RESULTS: Evidence of respiratory distress and a heart rate exceeding 120 beats per min were the two most sensitive indicators of gastrointestinal leak. Routine upper gastrointestinal contrast imaging detected only two of nine leaks (22%). CONCLUSION: Leak after LRYGB may be difficult to detect. Evidence of respiratory distress and tachycardia exceeding 120 beats per min may be the most useful clinical indicators of leak after laparoscopic Roux-en-Y gastric bypass.

Identification of a S-ribonuclease-binding protein in Petunia hybrida.

To investigate protein-protein interactions in gametophytic self-incompatibility, we used a yeast two-hybrid assay to identify proteins that could interact with the S-ribonuclease protein. These assays identified a pollen-expressed protein, which we have named PhSBP1, that appears to bind with a high degree of specificity to the Petunia hybrida S-ribonuclease. Although PhSBP1 activates reporter gene expression only when expressed in tandem with a S-RNAse bait protein, binding is not allele-specific. Sequence analysis demonstrated that PhSBP1 contained a C-terminal cysteine-rich region that includes a RING-HC domain. Because many RING-finger domain proteins appear to function as E3 ubiquitin ligases, our results suggest that ubiquitination and protein degradation may play a role in regulating self-incompatibility interactions. Together, these results suggest that PhSBPI may be a candidate for the recently proposed general inhibitor (RI) of self-incompatibility ribonucleases.

The S-ribonuclease gene of Petunia hybrida is expressed in nonstylar tissue, including immature anthers.

To determine the ability of isolated S-locus promoter sequences to direct organ-specific gene expression, we used microprojectile bombardment to introduce chimeric S-allele/beta-glucuronidase genes into different tissues of Petunia hybrida for transient expression. Histochemical staining showed that S-locus/beta-glucuronidase fusions were expressed in pistil, ovary, and petal tissue. No expression of the chimeric genes was detected in leaves or in mature pollen, either by histochemical staining or by fluorescence assays. RNA blot hybridization confirmed that low levels of S-locus mRNA accumulate in petals and ovaries in vivo. Analysis of the expression pattern of S-locus promoter deletions showed that sequences in the immediate vicinity of the TATA box were sufficient to confer qualitatively correct organ-specific expression of beta-glucuronidase. To further investigate the potential for S-ribonuclease expression in pollen, we used the polymerase chain reaction to amplify RNA accumulated in developing anthers. These assays demonstrated that mRNA for the S-ribonuclease accumulates to low levels in developing anthers several days prior to corolla opening and pollen anthesis. We discuss these results in light of current models of self-incompatibility.

Sequence variability and developmental expression of S-alleles in self-incompatible and pseudo-self-compatible petunia.

We investigated the structure and expression of three S-alleles of Petunia hybrida in self-incompatible varieties and in a pseudo-self-compatible line in which the self-incompatibility response is defective. Comparison of derived amino acid sequences from different gametophytic S-alleles revealed a pattern of sequence conservation and variability that was highly nonrandom. In self-incompatible varieties, petunia S-locus mRNA accumulates preferentially in styles during the transition from bud self-compatibility to self-incompatibility. S-Allele sequences homologous to the cloned S1 allele were present in a pseudo-self-compatible variety, and were expressed at levels indistinguishable from those observed in a self-incompatible line homozygous for the S1 allele. Taken together, our data indicate that (1) limited sequence differences may confer allelic specificity, (2) S-locus mRNAs accumulate in a precise organ-specific pattern during floral development, and (3) the ability to inhibit the growth of incompatible pollen tubes appears to require a threshold accumulation of the stylar gene product, along with the participation of as yet undefined pollen gene products.

Characterization of the glycinin gene family in soybean.

We characterized the structure, organization, and expression of genes that encode the soybean glycinins, a family of storage proteins synthesized exclusively in seeds during embryogenesis. Five genes encode the predominant glycinin subunits found in soybeans, and they have each been cloned, sequenced, and compared. The five genes have diverged into two subfamilies that are designated as Group-I and Group-II glycinin genes. Each glycinin gene contains four exons and three introns like genes that encode related proteins in other legumes. Two other genes have been identified and designated as "glycinin-related" because they hybridize weakly with the five glycinin genes. Although not yet characterized, glycinin-related genes could encode other glycinin subunit families whose members accumulate in minor amounts in seeds. The three Group-I glycinin genes are organized into two chromosomal domains, each about 45 kilobase pairs in length. The two domains have a high degree of homology, and contain at least five genes each that are expressed either in embryos or in mature plant leaves. Gel blot studies with embryo mRNA, as well as transcription studies with 32P-RNA synthesized in vitro from purified embryo nuclei, indicate that glycinin and glycinin-related genes become transcriptionally activated in a coordinated fashion early in embryogenesis, and are repressed coordinately late in seed development. In addition to transcriptional control processes, posttranscriptional events also are involved in regulating glycinin and glycinin-related mRNA levels during embryogenesis.

Light-stimulated increase of translatable mRNA for phosphoenolpyruvate carboxylase in leaves of maize.

Polyadenylated RNA has been isolated from maize leaves at various times during greening of etiolated seedlings and used to prime the wheat germ cell-free translation system. Levels of translatable messenger RNA for phosphoenolpyruvate carboxylase increase with the length of the illumination period. The pattern of the increase in translatable mRNA for the enzyme is similar to that of the increase in phosphoenolpyruvate carboxylase protein previously observed in intact tissues. Phosphoenolpyruvate carboxylase synthesized in vivo migrates as a doublet band on gradient sodium dodecyl sulfate-polyacrylamide gels. A similar doublet has been seen occasionally with the products of cell-free translation.

Evidence for Light-stimulated Synthesis of Phosphoenolpyruvate Carboxylase in Leaves of Maize.

Illumination (22,000 lumens per meter(2)) of etiolated maize plants for 80 hours brings about a 5-fold increase in phosphoenolpyruvate carboxylase activity per unit of protein. An increase in carboxylase protein and incorporation of [(35)S]methionine into the protein occurs simultaneously with the activity increase. In green plants, the level of phosphoenolpyruvate carboxylase protein and enzyme activity is dependent on the intensity of light during growth. These results are consistent with the conclusion that the activity increase results from light-stimulated de novo synthesis of phosphoenolypyruvate carboxylase protein.

Gray scale ultrasonography in the evaluation of hydrocephalus and associated abnormalities in infants.

Improved gray scale ultrasonic equipment has made two-dimensional echoencephalography a useful diagnostic tool in the examination of the infant brain for fluid-containing structures such as dilated ventricles and cysts. Eighteen infants have been examined to exclude hydrocephalus and other abnormalities. Ten had significantly dilated lateral ventricles and three of these were shown to have associated intracerebral abnormalities. Another infant was demonstrated to have an arachnoid cyst or subdural hygroma with normal size ventricles. Ten of the 11 abnormalities were confirmed by some other neuroradiologic study.