Improvements in life expectancy in adult type 2 diabetes patients in the last six decades.

AIM: To investigate the historical changes in survival with diabetes in adult type 2 diabetes patients. METHODS: We analyzed 9066 deaths, 54.2% males, registered at "I. Pavel" Bucharest Diabetes Centre, aged 40-64 years and deceased between 1943 and 2009. We split the analysis in three time periods according to year of death: 1943-1965, 1966-1988 and 1989-2009. RESULTS: The mean age at diabetes onset was 55.5 ± 6.2 years, with mean disease duration at death 12.7 ± 8.2 years and mean age at death 68.2 ± 8.7 years. The mean disease duration at death was 9.9 ± 7.3 years in 1943-1965 period, followed by a significant (p<0.001) rise to 12.2 ± 8.2 years in 1966-1988, and 14 ± 8.1 years (p < 0.001) in 1989-2009. There was a significant increase for coronary heart diseases and cancer and a significant decrease for infections and end-stage renal disease as causes of death. CONCLUSION: We found a significant increase in age at onset and survival with diabetes leading to a significant increase in age at death.

Identification of genes downstream of nodal in the Ciona intestinalis embryo.

Nodal, a growth factor belonging to the TGF-beta superfamily, is required for the formation of the neural tube in Ciona intestinalis. Previous studies have revealed many genes whose expression is controlled by Nodal in the Ciona embryo; however, all of them encode transcription factors and signaling molecules. In the present study, we identified five genes upregulated or downregulated by the overexpression of Nodal in embryos of C. intestinalis. The upregulated genes included those encoding type IV collagen 1/3/5, laminin-alpha5, and Prickle. The downregulated genes included those encoding glypican and delta1-protocadherln-like. Many of these genes were expressed in the neural plate at the late gastrula stage. The present study revealed candidate effector genes that directly regulate, in response to Nodal, the morphogenesis of the neural tube in Ciona intestinalis.

Improvements in life expectancy in type 1 diabetes patients in the last six decades.

AIMS: To investigate the survival with diabetes in patients treated with insulin from diagnosis. SUBJECTS AND METHODS: We analyzed 845 subjects, 55.9% males, registered at "I. Pavel" Bucharest Diabetes Centre, insulin-treated from diagnosis, aged <40 years and deceased between 1946 and 2005. We divided the subjects in two groups by age at diagnosis: group A <18 years and group B 18-39.99 years. We used 20 years time periods for year of death: 1946-1965, 1966-1985 and 1986-2005. RESULTS: The mean age at diabetes onset was 30.36+/-8.04 years, disease duration at death 20.98+/-11.62 years and age at death 51.34+/-14.37 years. The mean increase in survival with diabetes was 19.3 years for group A and 15.9 years for group B. There was a significant decrease in infections in both groups. The increase in coronary heart diseases and stroke is evident only in group B. CONCLUSIONS: We found no changes in age at onset, which combined with an increase in survival with diabetes lead to a significant increase in age at death over the six decades analyzed.

A novel method, digital genome scanning detects KRAS gene amplification in gastric cancers: involvement of overexpressed wild-type KRAS in downstream signaling and cancer cell growth.

BACKGROUND: Gastric cancer is the third most common malignancy affecting the general population worldwide. Aberrant activation of KRAS is a key factor in the development of many types of tumor, however, oncogenic mutations of KRAS are infrequent in gastric cancer. We have developed a novel quantitative method of analysis of DNA copy number, termed digital genome scanning (DGS), which is based on the enumeration of short restriction fragments, and does not involve PCR or hybridization. In the current study, we used DGS to survey copy-number alterations in gastric cancer cells. METHODS: DGS of gastric cancer cell lines was performed using the sequences of 5000 to 15000 restriction fragments. We screened 20 gastric cancer cell lines and 86 primary gastric tumors for KRAS amplification by quantitative PCR, and investigated KRAS amplification at the DNA, mRNA and protein levels by mutational analysis, real-time PCR, immunoblot analysis, GTP-RAS pull-down assay and immunohistochemical analysis. The effect of KRAS knock-down on the activation of p44/42 MAP kinase and AKT and on cell growth were examined by immunoblot and colorimetric assay, respectively. RESULTS: DGS analysis of the HSC45 gastric cancer cell line revealed the amplification of a 500-kb region on chromosome 12p12.1, which contains the KRAS gene locus. Amplification of the KRAS locus was detected in 15% (3/20) of gastric cancer cell lines (8-18-fold amplification) and 4.7% (4/86) of primary gastric tumors (8-50-fold amplification). KRAS mutations were identified in two of the three cell lines in which KRAS was amplified, but were not detected in any of the primary tumors. Overexpression of KRAS protein correlated directly with increased KRAS copy number. The level of GTP-bound KRAS was elevated following serum stimulation in cells with amplified wild-type KRAS, but not in cells with amplified mutant KRAS. Knock-down of KRAS in gastric cancer cells that carried amplified wild-type KRAS resulted in the inhibition of cell growth and suppression of p44/42 MAP kinase and AKT activity. CONCLUSION: Our study highlights the utility of DGS for identification of copy-number alterations. Using DGS, we identified KRAS as a gene that is amplified in human gastric cancer. We demonstrated that gene amplification likely forms the molecular basis of overactivation of KRAS in gastric cancer. Additional studies using a larger cohort of gastric cancer specimens are required to determine the diagnostic and therapeutic implications of KRAS amplification and overexpression.

Localization and characterization of γ-glutamyl cyclotransferase in cancer cells.

Using differential display analysis, we have identified a novel rat gene whose expression is increased during tumor progression in rat mammary carcinoma cell lines. This gene is an ortholog of the human chromosome 7 open reading frame 24 gene (C7orf24) and encodes a protein of 188 amino acids with no recognized protein domains. C7orf24 has been identified as γ-glutamyl cyclotransferase (GGCT), an important enzyme functioning in glutathione homeostasis. Our Northern and Western blot analyses revealed that the GGCT gene is expressed in various normal human and tumor tissues, as well as in cancer cell lines. Among the tumor tissues tested, lung tumor tissue expressed GGCT mRNA more strongly than normal lung tissue. The GGCT protein was found to be localized in the cytoplasmic region of cultured cells, where it forms a homodimer. Analysis of various deletion mutants of the GGCT protein revealed that the region containing amino acid residues 61-120 of the protein is required for its cytoplasmic localization. The comparison of the soft agar colony formation of HBL-100 cells stably expressing GGCT with that of control HBL-100 cells revealed that GGCT does not promote colony formation, suggesting that the role it plays in lung cancer cells is not related to tumorigenesis.

Gene expression profile during the life cycle of the urochordate Ciona intestinalis.

Recent whole-genome studies and in-depth expressed sequence tag (EST) analyses have identified most of the developmentally relevant genes in the urochordate, Ciona intestinalis. In this study, we made use of a large-scale oligo-DNA microarray to further investigate and identify genes with specific or correlated expression profiles, and we report global gene expression profiles for about 66% of all the C. intestinalis genes that are expressed during its life cycle. We succeeded in categorizing the data set into 5 large clusters and 49 sub-clusters based on the expression profile of each gene. This revealed the higher order of gene expression profiles during the developmental and aging stages. Furthermore, a combined analysis of microarray data with the EST database revealed the gene groups that were expressed at a specific stage or in a specific organ of the adult. This study provides insights into the complex structure of ascidian gene expression, identifies co-expressed gene groups and marker genes and makes predictions for the biological roles of many uncharacterized genes. This large-scale oligo-DNA microarray for C. intestinalis should facilitate the understanding of global gene expression and gene networks during the development and aging of a basal chordate.

cDNA microarray analyses reveal candidate marker genes for the detection of ascidian disease in Korea.

A serious disease of the ascidian Halocynthia roretzi has been spread extensively among Korean aquaculture sites. To reveal the cause of the disease and establish a monitoring system for it, we constructed a cDNA microarray spotted with 2,688 cDNAs derived from H. roretzi hemocyte cDNA libraries to detect genes differentially expressed in hemocytes between diseased and non-diseased ascidians. We detected 21 genes showing increased expression and 16 genes showing decreased expression in hemocytes from diseased ascidians compared with those from non-diseased ascidians. RT-PCR analyses confirmed that the expression levels of genes encoding astacin, lysozyme, ribosomal protein PO, and ubiquitin-ribosomal protein L40e fusion protein were increased in hemocytes from diseased ascidians, while those of genes encoding HSP40, HSP70, fibronectin, carboxypeptidase and lactate dehydrogenase were decreased. These genes were expressed not only in hemocytes but also in various other tissues in ascidians. Furthermore, the expression of glutathione-S transferase omega, which is known to be up-regulated in H. roretzi hemocytes during inflammatory responses, was strongly increased in hemocytes from diseased ascidians. These gene expression profiles suggest that immune and inflammatory reactions occur in the hemocytes of diseased ascidians. These genes will be good markers for detecting and monitoring this disease of ascidians in Korean aquaculture sites.

A pathophysiological approach to metabolic syndrome using factor analysis in an adult Romanian population.

The aim of the study was to examine the role of insulin resistance in etiopathogenesis of metabolic syndrome in an adult Romanian population using exploratory factor analysis. We analyzed 228 non-diabetic subjects randomized in respect to the age and sex distribution of the general population. For each patient, age, sex, body mass index (BMI), systolic and diastolic blood pressure (SBP, DBP), HDL-cholesterol (HDL), plasma triglycerides (TG), fasting plasma glucose (FPG) and fasting insulin were obtained. Factor analysis was performed using principal component analysis, with Varimax rotation of the major determinants of metabolic syndrome. Mean age was 48.9 +/- 12.7 years; 107 (46.9%) were men and 121 (53.1%) women. We found three major factors, which are correlated with metabolic syndrome and may explain its variance. Factor 1 comprises SBP and DBP in men and SBP, DBP and BMI in women. Factor 2 comprises BMI, HDL, TG and FPG in men and BMI, TG and FPG in women. Factor 3 comprises fasting insulin in men and fasting insulin, TG and HDL in women. The finding of more than one factor suggests that insulin resistance is not the only pathophysiological mechanism involved. These factors appear to work independently of each other in men, but they intersect in women, suggesting that the pathophysiology of metabolic syndrome may be different in women compared with men.