Thymidylate synthase expression in gastroenteropancreatic and pulmonary neuroendocrine tumors

PURPOSE: The predictive role of the quantification of thymidylate synthase (TS) in tumors treated with antifolate drugs, such as 5-fluorouracil (5-FU), has been extensively reported in a variety of human tumors. Neuroendocrine tumors (NET) represent potential targets of antifolate agents, but no data on TS expression level in these tumors are currently available. EXPERIMENTAL DESIGN: A series of 116 NETs were collected, including 58 gastroenteropancreatic (GEP) and 58 lung NETs. In 24 well-differentiated GEP neuroendocrine carcinomas (WD-NEC), a 5-FU-based treatment was given. Total RNA was extracted from microdissected paraffin blocks. TS mRNA quantification was done by real-time PCR, whereas protein expression was evaluated by immunohistochemistry. RESULTS: By means of both quantification by real-time PCR and immunohistochemistry, a higher TS expression in pulmonary small cell lung cancer and large cell NEC compared with typical and atypical carcinoids was observed (P < 0.01). Similarly, in GEP tumors, a higher TS expression in poorly differentiated carcinomas than both WD-NEC and benign tumors (P < 0.01) was found. In patients with WD-NEC treated with 5-FU, high TS mRNA levels were associated with shorter time to progression (P = 0.002) and overall survival (P = 0.04). This negative prognostic role was confirmed in multivariate analysis adjusting for major prognostic variables (P = 0.01). No association between TS mRNA and survival was observed in WD-NEC patients not receiving 5-FU. CONCLUSIONS: This study, for the first time, (a) reports the differential TS expression in the spectrum of NETs and (b) indicates TS as a possible predictive marker of treatment efficacy in WD-NEC patients treated with 5-FU.     

Molecular and Histological Changes in Post-Treatment Biopsies of Non-Squamous Non-Small Cell Lung Cancer: A Retrospective Study

Background

Recently, in advanced non-small cell lung cancer (NSCLC), standard chemotherapy was flanked by biological agents directed against genomic abnormalities, including EGFR and ALK alterations, that significantly improved patient outcome. Despite these achievements, tumour progression almost always occurs and a reassessment of the tumour genetic profile may contribute to modulating the therapeutic regimen. Resampling may provide tissue for additional tests to detect acquired resistance and/or new genetic alterations, but the currently available information is limited.

Patients and Methods

Histological and genetic reassessments of biopsy or surgical tissue samples from 50 non-squamous NSCLC patients before and after at least one systemic treatment were performed. EGFR, KRAS, BRAF, PIK3CA and HER2 mutations were sequenced, p.T790M was identified with real-time PCR, and ALK and MET genomic alterations by fluorescence in situ hybridization.

Results

Overall in baseline biopsies, 37/50 (74 %) tumours had genetic alterations, either single (52 %) or multiple (22 %). Among them, 16 were EGFR mutations and 6 ALK rearrangements. In the second tissue sampling, 54 % of cases had additional genomic changes, including newly acquired alterations (81 %) or losses (18 %). The commonest changes were MET amplification and p.T790M mutation. One case had a histological shift from adenocarcinoma to small cell carcinoma.

Conclusions

The remarkable number of molecular changes following systemic therapy and the genetic complexity of some cases underline the value of histological and molecular re-evaluation of lung cancer to tailor the most appropriate therapy during disease progression.

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The over-expression of cell migratory genes in alveolar rhabdomyosarcoma could contribute to metastatic spread

Alveolar (ARMS) and Embryonal (ERMS) rhabdomyosarcoma differ in their response to current treatments. The ARMS subtype has a less favourable prognosis and often presents with widespread metastases, while the less metastatic ERMS has a 5 year survival rate of more than 80 %. In this study we investigate gene expression differences that could contribute to the high frequency of metastasis in ARMS. Microarray analysis identified significant differences in DNA repair, cell cycle and cell migration between the two RMS subtypes. Two genes up regulated in ARMS and involved in cell migration; the engulfment and cell motility gene 1 (ELMO1) and NEL-like 1 gene (NELL1) were selected for further investigation. Over-expression of ELMO1 significantly increased cell invasion from 24.70 ± 7 % to 93 ± 5.4 % in primary myoblasts and from 29.43 ± 2.1 % to 87.33 ± 4.1 % in the ERMS cell line RD. siRNA knockout of ELMO1 in the ARMS cell line RH30 significantly reduced cell invasion from 88.2 ± 3.8 % to 35.2 ± 2.5 %. Over-expression of NELL1 significantly increased myoblast invasion from 23.6 ± 6.9 % to 100 ± 0.1 %, but had no effect on invasion of the ERMS cell line RD. These findings suggest that ELMO1 may play a key role in ARMS metastasis. NELL1 increased invasion in primary myoblasts, but other factors required for it to enhance motility were not present in the RD ERMS cell line. Impairing ELMO1 function by pharmacological or siRNA knockdown could be a highly effective approach to reduce the metastatic spread of RMS.     

Nonsymptomatic changes in magnetic resonansce imaging of spine among airforce candidates]

BACKGROUND: Problems of low back and neck pain are were important for persons, who decided to participate in sport or to educate and work in jobs requiring high accelerations and loads. The aim of the study was evaluate a structure the whole spine on magnetic resonansce imaging among airforce candidates. MATERIAL AND METHODS: MRI of 98 young, healthy persons, with no abnormalities in structure and function of musclosceletal system, were analysed. RESULTS: Analysed material revealed dehydratation of intervertebral discs L4-L5 and L5-S1 among 23 persons (23.5%). In this group 21 persons (22%) had protrusions, while Schmorl nodules were present among 14 (14.7%) persons. Only one persons had protrusion in cervical spine. Candidates who had changes in MRI were excluded from futher qualifying selection. An exeption was Schmorl nodules in thoracal spine due to good stabilisation of this part of spine gained by rips. CONCLUSION: (1) Intervertebral disc degeneration without clinical symptoms were found among 23% examined persons and starts in second decade of life. (2) Aboved mentioned changes are a serius problem for airforce candidates. (3) Airforce candidates, who have degenerative changes in lumbar and cervical spine on MRI findings should not be exepted to futher qualifying selection.     

Ghrelin secretion is inhibited by glucose load and insulin-induced hypoglycaemia but unaffected by glucagon and arginine in humans

OBJECTIVE: Circulating ghrelin levels are increased by fasting and decreased by feeding, glucose load, insulin and somatostatin. Whether hyperglycaemia and insulin directly inhibit ghrelin secretion still remains matter of debate. The aim of the present study was therefore to investigate further the regulatory effects of glucose and insulin on ghrelin secretion. DESIGN AND SUBJECTS: We studied the effects of glucose [oral glucose tolerance test (OGTT) 100 g orally], insulin-induced hypoglycaemia [ITT, 0.1 IU/kg insulin intravenously (i.v.)], glucagon (1 mg i.v.), arginine (0.5 mg/kg i.v.) and saline on ghrelin, GH, insulin, glucose and glucagon levels in six normal subjects. MEASUREMENTS: In all the sessions, blood samples were collected every 15 min from 0 up to + 120 min. Ghrelin, GH, insulin, glucagon and glucose levels were assayed at each time point. RESULTS: OGTT increased (P < 0.01) glucose and insulin while decreasing (P < 0.01) GH and ghrelin levels. ITT increased (P < 0.01) GH but decreased (P < 0.01) ghrelin levels. Glucagon increased (P < 0.01) glucose and insulin without modifying GH and ghrelin. Arginine increased (P < 0.01) GH, insulin, glucagon and glucose (P < 0.05) but did not affect ghrelin secretion. CONCLUSIONS: Ghrelin secretion in humans is inhibited by OGTT-induced hyperglycaemia and ITT but not by glucagon and arginine, two substances able to increase insulin and glucose levels. These findings question the assumption that glucose and insulin directly regulate ghrelin secretion. On the other hand, ghrelin secretion is not associated with the GH response to ITT or arginine, indicating that the somatotroph response to these stimuli is unlikely to be mediated by ghrelin.     

Evaluation of various methods treatment of proximal humerus fractures

Background. Most of the proximal humerus fractures are osteoporotic and concern elderly patients. In the literature long-term results of treatment of proximal humerus fractures are poor or fair. Objective. The objective of the paper was to present various methods of treatment - conservative and surgical. Material and method. A group of 138 fractures of proximal humerus fractures treated during years 1988-1999 was analysed. 84 (63%) fractures were followed up for a period ranging from 18 months to 10 years. Results were estimated by Neer's criteria. Results. The worst results in conservative treatment were obtained when Desault plaster cast was used (47% poor and fair); the best when sling was used (75% very good and good). Among the surgical methods the best results were observed thanks to fixation with threaded Kirschner wires (86% very good and good); the worst when screw or plate and screw fixation was used (60% poor). Conclusions. 1. For two- and three-part fractures with dislocation in which proper heeling augurs well, the right method of fixation is stabilisation with threaded Kirschner wires. 2. Treatment to be chosen for fractures without dislocation is the sling and early rehabilitation (1-2 weeks after trauma), whereas treatment in Desault plaster cast should be regarded as a mistake. 3. In elderly patients with four-part fractures, haemiarthroplasty as a primary operation should be considered.