The Early Growth Response Gene EGR-1 Behaves as a Suppressor Gene That Is Down-Regulated Independent of ARF/Mdm2 but not p53 Alterations in Fresh Human Gliomas.

EGR-1 is an immediate early gene with diverse functions that include the suppression of growth. EGR-1 is down-regulated many cancer cell types, suggesting a tumor suppressor role, and may critically involve the p53 pathway. The aim of this work was to measure the expression of EGR-1 and the p16/INK4a/ARF-Mdm2-p53 pathway status in fresh human gliomas. Thirty-one human gliomas with different grades of malignancy were investigated for Egr-1 mRNA and the protein expression, frequency, and spectrum of p53 gene mutations, mdm2 gene amplification, and p16/INK4a/ARF allele loss. The amplification of Mdm2 and the deletion of the p16/INK4a gene was found in 3 and 5 cases, respectively, whereas mutations of p53, including two novel mutations, were observed in 10 other cases. The three types of changes occurred strictly mutually exclusively, emphasizing that these genes operate in a common pathway critical to glioma progression. EGR-1 mRNA was significantly down-regulated in astrocytomas (14.7 +/- 5.1%) and in glioblastomas (33.6 +/- 10.0%) versus normal brain. Overall, EGR-1 mRNA was strongly suppressed (average, 15.2 +/- 13.9%) in 27 of 31 cases (87%), independent of changes in p16/INK4a/ARF and Mdm2; whereas 4 of 31 cases with residual EGR-1 expression as well as the highest EGR-1 variance segregated with p53 mutations. Immunohistochemical analyses confirmed the suppression of EGR-1 protein. These results indicate that EGR-1 is commonly suppressed in gliomas independent of p16/INK4a/ARF and Mdm2 and that suppression is less crucial in tumors bearing p53 mutations, and these results implicate an EGR-1 growth regulatory mechanism as a target of inactivation during tumor progression

Prognostic value of splenectomy and lymph-node dissection during gastric cancer resection

Gastric carcinoma is the second most common cause of digestive tumour-related death in Europe, North America and Asia. Today, the gold standard of treatment is still surgery, but outcomes to date are unsatisfactory. The Japanese Society for Research in Gastric Cancer (JSRGC) recommends the routine execution of splenectomy during gastrectomy. This recommendation is contested by western Authors because of increased morbidity and mortality without any real advantage in terms of survival. Patients treated for gastric cancer in our department between 1993 and 2002 were selected for this study. The 132 patients recruited were divided into two groups: a) those treated for gastric cancer without splenectomy; and b) those in whom splenectomy was performed in conjunction with gastrectomy. We analysed: the extent of lymph node dissection, the execution of the splenectomy, and the prognostic importance of factors relating to the patient, tumour and surgeon. Splenectomy was not associated with any increase in morbidity. Complications, especially of the septic type, and perioperative mortality were similar in both groups, and the same was true of survival at both 3 and 5 years. In our opinion, splenectomy should not be routinely combined with surgery for gastric cancer but could be considered for T3-T4 neoplasms or those localized in the upper two thirds of the stomach

Navigated, soft tissue-guided total knee arthroplasty restores the distal femoral joint line orientation in a modified mechanically aligned technique

Purpose: To investigate the femoral component alignment in patients undergoing soft tissue-guided, navigated total knee arthroplasty (TKA). It was hypothesized that with a mechanically aligned tibial component, the soft tissues tensioned and symmetric medial and lateral gaps in flexion/extension, the femoral component would be aligned to the preoperative distal femoral joint line, as measured on knee radiographs. Methods: Between 2015 and 2017, 77 patients (78 knees) underwent navigated soft tissue-guided TKA at a single centre. Pre and postoperative radiographs were collected and varus knees were taken into account. The tibial cut was performed with navigation in neutral alignment. The femoral cuts were adjusted based on tensioned soft tissues, aiming for equal medial and lateral gaps in flexion and extension. The Knee Injury and Osteoarthritis Outcome Score (KOOS) and Knee Society Score (KSS) were collected pre and postoperatively as a secondary outcome measure. Results: A total of 58 TKAs on varus knees were assessed. On average, the femoral component was placed at 1.7° (SD 1.7) varus in the coronal plane. The comparison between the radiographic native distal femoral alignment and the orientation of the femoral component coronal cut demonstrated a statistically significant (p < 0.0001), linear inverse relationship (r = 0.5). Satisfactory knee function and excellent pain remission were demonstrated by KOOS and KSS scores at a mean of 2.8 years (SD 0.5) follow-up. One TKA was revised, resulting in a 98.3% survivorship at three years. Conclusion: The proposed soft tissue-guided, navigated technique, aiming to preserve the integrity of the ligaments and a neutrally aligned tibial cut, provided a joint line respecting femoral coronal cut and encouraging short-term clinical results. Level of evidence: III

The oncoprotein Myc controls the phosphorylation of S6 kinase and AKT through protein phosphatase 2A

This study focuses on the effects of Myc oncoprotein on the translational apparatus of the cell. Translation is an energy consuming process that involves a large number of accessory factors. The production of components of the protein synthesis machinery can be regulated at the transcriptional level by specific factors. It has been shown that the product of the oncogene Myc, a transcription factor frequently activated in cancer, can control translational activity through an increase in the transcription of the eIF4F complex components (eIF4E, eIF4AI, and eIF4GI). However, additional effects at the posttranslational level have also been described. For instance, it has been shown that Myc upregulation can induce mammalian target of rapamycin (mTOR)-dependent 4E-binding protein 1 (4E-BP1) hyperphosphorylation. We induced overexpression or inhibition of Myc through transfection of complementary DNA constructs or specific small interfering RNA in PC3 (prostate carcinoma) and HeLa (cervical carcinoma) cells. We have observed that overexpression of Myc causes an increase in 4E-BP1 phosphorylation and activation of protein synthesis. Unexpectedly, we detected a parallel decrease in the phosphorylation level of S6 kinase (in PC3 and HeLa) and AKT (in HeLa). We report evidence that these changes are mediated by an increase in protein phosphatase 2A activity

Inhibition of MAPK activity ,cell proliferation and anchorage-independent growth by N-ras antisense in an N-ras transformed human cell line.

Mammalian ras genes encode a family of plasma membrane-bound proteins that function as intermediates in signal transduction pathways involved in cell growth and differentiation. Ras oncogene is frequently involved in neoplastic transformation of different cellular histotypes. In this study, we tested the ability of antisense oligodeoxyribonucleotides (AS-ODN) that have mixed phosphodiester/phosphorothioate backbone, targeted against human N-Ras, to inhibit N-ras gene expression and to specifically interfere with the Ras-dependent activity of mitogen-activated protein kinase (MAPK) in two human cell lines carrying an endogenous N-ras mutated allele at codon 61. Three AS-ODN that inhibit basal MAPK activity have been identified. Moreover, AS-ODN treatment resulted in potent antiproliferative effects in cell culture and great inhibition of N-ras mRNA levels in one of two cell lines. These studies suggest that antisense molecules, targeted against N-Ras, could be of considerable value as a tool to study the N-Ras-specific transduction pathway

Cell-mediated response in cattle experimentally infected with bluetongue virus serotype 2

Cell-mediated immunity in cattle infected with bluetongue virus serotype 2 was examined using the 3-(4,5, dimethylthiazol- 2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) lymphocyte proliferation assay and the enzyme-linked immunosorbent assay (ELISA) kit for gamma-interferon quantification in serum. Although infection induced the production of neutralising antibodies, no significant statistical differences were observed between the infected and the control animals when tested with the MTT assay. Constant levels of gamma-interferon were detected in the serum infected animals during the trial but again no significant statistical differences were recorded. The results of the study are discussed.[...