Influence of glioblastoma contact with the subventricular zone on survival and recurrence patterns.

BACKGROUND: There is growing evidence that the subventricular zone (SVZ) may be involved in both the initiation and progression of glioblastoma (GB). We aimed to assess tumor proximity to the SVZ as a potential prognostic factor in GB. METHOD: Retrospective study of 133 patients diagnosed with primary GB who underwent surgery followed by temozolomide-based chemoradiation between 2010 and 2016. All lesions were classified according to their anatomic relation with the SVZ. We determined the effect of tumor contact with the SVZ on progression-free survival (PFS), overall survival (OS), type, and patterns of recurrence. RESULTS: At a median follow-up of 18.6 months (95% CI 15.9-21.2), PFS and OS were 7.5 (95% CI 6.7-8.3) and 13.9 (95% CI 10.9-16.9) months, respectively. On the univariate analyses, initial contact with the SVZ was a factor for poor prognosis for both PFS (6.1 vs. 8.7 months; p = 0.006) and OS (10.6 vs. 17.9 months; p = 0.037). On the multivariate analysis, tumor contact with the SVZ remained statistically significant for PFS, but not OS. Patients with SVZ-contacting tumors presented a higher rate of aggressive clinical progression (30.9% vs. 11.3%; p = 0.007) and contralateral relapse patterns (23.4% vs. 9.1%; p = 0.048). CONCLUSIONS: Our results suggest that glioblastoma contact with the SVZ appears to be an independent prognostic factor for poor PFS. The presence of an SVZ-contacting tumor was associated with more aggressive recurrences and a higher rate of contralateral relapses. These findings suggest that this variable may be a new prognostic factor in glioblastoma.

Assessment of neurocognitive decline in cancer patients, except brain cancer, under long-term treatment with bevacizumab.

PURPOSE: We performed a cross-sectional study of neurocognitive function in non-brain cancer patients treated with long-term bevacizumab. METHODS/PATIENTS: From 2015 to 2017, we included patients with different types of cancer treated with bevacizumab with or without chemotherapy (BEV; N = 20) or only chemotherapy (ChT; N = 19) for at least 34 weeks, patients who received non-brain radiotherapy (RxT; N = 19), and healthy controls (HC; N = 19) were assessed once at week 34 of treatment (BEV and ChT) or at completion of radiotherapy. Neurocognition was evaluated with the Hopkins Verbal Learning Test-Revised (HVLT-R) total and delayed recall, the Trail Making Test A and B, and the Controlled Oral Word Association Test in the four groups. Non-parametric tests were used to assess differences between groups. RESULTS: The BEV, ChT, and RxT groups scored significantly lower than the HC group on all tests and especially on the HVLT-R total recall. In no case were the mean scores of the BEV group significantly lower than those of the ChT or RxT groups. CONCLUSIONS: Neurocognitive impairment was seen even in patients treated with local non-brain radiotherapy. Treatment with bevacizumab for a long period of time does not seem to worsen neurocognitive function to a greater extent than chemotherapy.

SEOM clinical guideline on unknown primary cancer (2017).

Cancer of unknown primary site is a histologically confirmed cancer that manifests in advanced stage, with no identifiable primary site following standard diagnostic procedures. Patients are initially categorized based on the findings of the initial biopsy: adenocarcinoma, squamous-cell carcinoma, neuroendocrine carcinoma, and poorly differentiated carcinoma. Appropriate patient management requires understanding several clinical and pathological features that aid in identifying several subsets of patients with more responsive tumors.

Delay in starting radiotherapy due to neoadjuvant therapy does not worsen survival in unresected glioblastoma patients.

PURPOSE: We retrospectively examined the potential effect on overall survival (OS) of delaying radiotherapy to administer neoadjuvant therapy in unresected glioblastoma patients. PATIENTS AND METHODS: We compared OS in 119 patients receiving neoadjuvant therapy followed by standard treatment (NA group) and 96 patients receiving standard treatment without neoadjuvant therapy (NoNA group). The MaxStat package of R identified the optimal cut-off point for waiting time to radiotherapy. RESULTS: OS was similar in the NA and NoNA groups. Median waiting time to radiotherapy after surgery was 13 weeks for the NA group and 4.2 weeks for the NoNA group. The longest OS was attained by patients who started radiotherapy after 12 weeks and the shortest by patients who started radiotherapy within 4 weeks (12.3 vs 6.6 months) (P = 0.05). OS was 6.6 months for patients who started radiotherapy before the optimal cutoff of 6.43 weeks and 19.1 months for those who started after this time (P = 0.005). Patients who completed radiotherapy had longer OS than those who did not, in all 215 patients and in the NA and NoNA groups (P = 0.000). In several multivariate analyses, completing radiotherapy was a universally favorable prognostic factor, while neoadjuvant therapy was never identified as a negative prognostic factor. CONCLUSION: In our series of unresected patients receiving neoadjuvant treatment, in spite of the delay in starting radiotherapy, OS was not inferior to that of a similar group of patients with no delay in starting radiotherapy.

Results of a multicenter survey showing interindividual variability among neurosurgeons when deciding on the radicality of surgical resection in glioblastoma highlight the need for more objective guidelines.

PURPOSE: We assessed agreement among neurosurgeons on surgical approaches to individual glioblastoma patients and between their approach and those recommended by the topographical staging system described by Shinoda. METHODS: Five neurosurgeons were provided with pre-surgical MRIs of 76 patients. They selected the surgical approach [biopsy, partial resection, or gross total resection (GTR)] that they would recommend for each patient. They were blinded to each other's response and they were told that patients were younger than 50 years old and without symptoms. Three neuroradiologists classified each case according to the Shinoda staging system. RESULTS: Biopsy was recommended in 35.5-82.9%, partial resection in 6.6-32.9%, and GTR in 3.9-31.6% of cases. Agreement among their responses was fair (global kappa = 0.28). Nineteen patients were classified as stage I, 14 as stage II, and 43 as stage III. Agreement between the neurosurgeons and the recommendations of the staging system was poor for stage I (kappa = 0.14) and stage II (kappa = 0.02) and fair for stage III patients (kappa = 0.29). An individual analysis revealed that in contrast to the Shinoda system, neurosurgeons took into account T2/FLAIR sequences and gave greater weight to the involvement of eloquent areas. CONCLUSIONS: The surgical approach to glioblastoma is highly variable. A staging system could be used to examine the impact of extent of resection, monitor post-operative complications, and stratify patients in clinical trials. Our findings suggest that the Shinoda staging system could be improved by including T2/FLAIR sequences and a more adequate weighting of eloquent areas.

Comparison of two types of inocula during acclimation and stable operation for nitrophenol biodegradation in an anaerobic-aerobic SBR.

This work presents a comparison of two inocula used for the acclimation of two anaerobic-aerobic sequencing batch bioreactors used for toxic wastewater treatment. The bioreactors were acclimated with different types of sludge: one coming from an anaerobic wastewater treatment plant and the other one from a conventional aerobic activated sludge plant. The model toxic compound was p-nitrophenol, which is reduced to p-aminophenol during the initial anaerobic phase of the reaction, and later mineralized during a posterior aerated reaction phase. Biodegradation of the compounds was monitored using UV/Vis spectrophotometry. After acclimation stabilization of the sludge and of the process was also monitored. Results show that there is no significant difference in acclimation times and stability of the process between the two employed inocula, and thus an originally anaerobic inoculum presents no apparent advantage over a more easily accessible aerobic one.

Lipid-dependent proliferation of pancreatic cancer cell lines.

Capan 1, a human pancreatic cancer cell line, is routinely grown in 10% fetal calf serum (FCS). In order to characterize the factors needed for its proliferation, FCS was replaced by a synthetic serum (Ultroser G). For Capan 1 proliferation we found that Ultroser G was as efficient as FCS. A subfraction of Ultroser G containing insulin, transferrin, and lipids was found to be responsible for that response since a combination of these compounds reproduced the growth observed with 10% FCS. Lipids stimulated cell proliferation even in the absence of other factors. Other human (MIA PaCa 2, AsPC1, Panc 1) or rat (AR4-2J) pancreatic cancer cell lines tested proliferated in the reconstituted medium containing insulin (100 ng/ml), transferrin (100 micrograms/ml), fatty acid-free albumin (1 mg/ml), and bovine serum lipids (0.7%), as in 10% FCS. Furthermore, the growth of nonpancreatic cell lines (HT29, A431, CREF) was not enhanced by lipids. Lipoproteins were found to be involved in the mitogenic response of pancreatic cells to lipids, whereas phosphatidylcholine and fatty acids were either inefficient or inhibitors (MIA PaCa2 and AR4-2J). Alkaline phosphatase and amylase content, differentiation markers for Capan 1 and AR4-2J cells, respectively, were not modified by the reconstituted medium. These data suggest that lipids, insulin, and transferrin are the essential factors for the proliferation of pancreatic cancer cell lines, reproducing the growth effect of 10% FCS. Moreover, in the absence of most of the seric growth factors, pancreatic cells remained differentiated.

Morphological and biological modifications induced in a rat pancreatic acinar cancer cell line (AR4-2J) by unscheduled expression of basic fibroblast growth factors.

The role of the different basic fibroblast growth factor (bFGF) forms on the regulation of pancreatic acinar cancer cells was analyzed on the rat cell line AR4-2J. This cell line expresses bFGF receptors but does not produce bFGF. AR4-2J cells were retrovirally transfected with the wild type or with point-mutated bFGF complementary DNAs in order to obtain the expression of all the bFGF forms (clone A4), or of that of the M(r) 22,500 form (clone A3), or of that of the M(r) 18,000 form (clone A5). Each clone was less tumorigenic in nude mice than AR4-2J cells. In culture, only the coexpression of all the bFGF forms modified cell morphology (fibroblast-like) and secretory enzyme synthesis (about a 20-fold decrease of amylase and lipase). Cells expressing the high molecular weight bFGF (A3 and A4) were able to grow in serum-free medium. As for AR4-2J, exogenously added bFGF still exerted mitogenic effects on the bFGF-producing cells. These results suggest that pancreatic acinar cancer cells may respond to endogenous bFGF; furthermore, they seem very sensitive to the coexpression of the different bFGF forms which is often described in cancer cells.

SEOM Clinical Guideline for gastrointestinal sarcomas (GIST) (2016).

Gastrointestinal stromal tumors (GISTs) are the most common mesenchymal neoplasms of the digestive tract, and this disease has served as a paradigmatic model for successful rational development of targeted therapies. The introduction of tyrosine kinase inhibitors with activity against KIT/PDGFRA in both localized and advanced stages has remarkably improved the survival in a disease formerly deemed resistant to all systemic therapies. The Spanish Society of Medical Oncology (SEOM) guidelines provide a multidisciplinary and updated consensus for the diagnosis and treatment of GIST patients. We strongly encourage that the managing of these patients should be performed within multidisciplinary teams in reference centers.

Oxyntomodulin and related peptides control somatostatin secretion in RIN T3 cells.

We studied the effects of oxyntomodulin (OXM), of its C-terminal (19-37) fragment (OXM (19-37)) and of glucagon (GLU) on somatostatin release, cyclic AMP accumulation and inositol phosphate turnover in somatostatin-secreting RIN T3 cells in culture. Rapid changes in cellular free Ca2+ were also measured using fura-2. Carbachol was used as a control test agent for the parameters involving the inositol phosphate/Ca2+ cascade. OXM, GLU and OXM (19-37) were all able to stimulate somatostatin release with relative ED50 of approx. 1, 22 and 45, respectively. OXM and GLU stimulated cyclic AMP levels with relative ED50 of approx. 1 and 30, respectively, whereas OXM (19-37) was totally ineffective on this parameter. In contrast to carbachol, none of the peptides significantly modified the inositol phosphate turnover or induced rapid changes in cellular free Ca2+. We conclude that the RIN T3 cells contain a receptor-cyclic AMP system similar to that found in gastric mucosa and that this system is linked to somatostatin release. Another receptor-second messenger mechanism linked to somatostatin release is triggered by the (19-37) fragment. This mechanism is not the inositol phosphate/Ca2+ cascade triggered in the same cells by cholinergic agents.

Expression of two FGF-2 isoforms in pancreatic acinar cells (AR4-2J). Intracellular localization and role in the regulation of the extracellular matrix biosynthesis.

Fibroblast growth factor (FGF-2) is a multifunctional growth factor. In cells producing this factor, FGF-2 is synthesized as different molecular weight isoforms lacking the signal peptide sequence for secretion. All forms are highly concentrated in cells. The presence of a nuclearization signal sequence in some isoforms suggests the involvement of these isoforms in cell functions bypassing the cell surface receptors. Our aims were to better define the intracellular localizations of the FGF-2 isoforms by immunocytochemistry and confocal microscopy and to analyze whether these isoforms were involved in the expression of extracellular matrix (ECM) components. We chose the pancreatic acinar cell line AR4-2J since it does not synthesize FGF-2. These cells were retrovirally transfected by point-mutated FGF-2 cDNAs. The cell lines obtained produced either the 18 kDa form (A5 cells) or the 22.5 kDa form (A3 cells). In A5 cells, the 18 kDa form was found in the cytoplasm, on the cell surface reflecting its secretion, and in the nucleoli. Parental AR4-2J cells treated with exogenous FGF-2 exhibited identical localizations, suggesting that in A5 cells the 18 kDa form followed the same translocation pathways than the exogenous FGF-2. By contrast, in A3 cells the 22.5 kDa form was predominantly localized in the nucleoplasm but was undetectable on the cell surface, suggesting its direct translocation to the nucleus. Northern and Western blot analysis showed that cells expressing the high molecular weight form exhibited a decrease of laminin B1 protein level and mRNA stability. In contrast, collagen IV and fibronectin expressions were unmodified either in FGF-2-transfected cells or in parental cells treated by exogenous FGF-2. Thus, these data indicate that: 1) 18 and 22.5 kDa FGF-2 are preferentially localized in different nuclear compartments and 2) the high molecular weight form plays a role on the expression of some ECM components.

Analysis of somatostatin peptides produced by an endocrine pancreatic cell line.

Biological active forms of somatostatin are produced by cleavage of large precursors. If the sequence of the pre-proform of somatostatin has been deduced from cDNA structure in several species, little is known about the processing of these large precursors. For this purpose, the analysis of immunoreactive components secreted by the R.I.N. cell line was investigated. After selection of a cell population and culture conditions providing the optimal production of these peptides, analysis of their molecular forms was done by molecular gel filtration. The results show that mainly pro-forms accumulate in the culture medium while besides the pre-proform the smaller immunoreactive species behaving like S-28 and S-14 were found in cell extracts. Incorporation studies in serum free medium showed rapid formation of an intermediate compound eluted at 1.87 V0.

Evidence for specific involvement of 5-HT1A and 5-HT2A/C receptors in the expression of patterns of spontaneous motor activity of the rat.

The 5-HT1A and the 5-HT2A/C receptor agonists 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT) (0.006-0.4 mg kg-1 s.c.) and (+/-)-1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane (DOI) (0.05-4.0 mg kg-1 s.c.), respectively, produced a similar stereotyped forward locomotion in rats, although the intensity of the behavioral change was considerably less with DOI. The stereotyped forward locomotion was accompanied by a slight decrease in total activity, suppression of rearing behavior and an increased activity in the periphery of the open-field arena. In support of receptor specificity, the effects of 8-OH-DPAT and DOI could be antagonised by pretreatment with the 5-HT1A/B and the 5-HT2A/C receptor antagonists (-)-pindolol (2 mg kg-1 s.c.) and ritanserin (2 mg kg-1 s.c.), respectively. In addition, (-)-pindolol, but not the selective beta-adrenoceptor antagonist betaxolol, markedly enhanced the behavioral effects produced by DOI. The nature of these specific actions and interactions in terms of pre- and post-synaptic serotonergic mechanisms remains an important question.

Lipid content of human and rat pancreas.

We analyzed the lipid composition of the human pancreas and performed a parallel study on rat pancreas. Some precautions were taken in order to keep the secretory zymogens as inactive precursors in both tissues. The lipid content of the human pancreas corresponded to 5.5% of the tissue wet weight, lower than that found in pancreas of two-month-old Wistar rats (10%). In man, triglycerides and phospholipids were found at comparable levels, respectively, 37 and 30 mg/g of pancreas wet weight, not far from the values of the rat pancreas. In human pancreas, phosphatidylcholines and lysophosphatidylcholines represented about 40% of the total phospholipid fraction, phosphatidylethanolamines and lysophosphatidylethanolamines 21%, and phosphatidylserines and -inositols were found equally represented with 7.5%. The total cholesterol content accounted for about 4.5% of the total lipids; only 30% was esterified. By comparison, in rat, total cholesterol represented 3.3% of lipids and 90% was esterified. The phospholipids in human pancreas contained high amounts of saturated fatty acids (92%) mainly stearic and palmitic, whereas triglycerides contained equal amounts of saturated and unsaturated fatty acids, principally represented by oleic and palmitic acids. In rats the phospholipids contained only 63% saturated fatty acids (palmitic and stearic) and triglycerides contained 61% unsaturated fatty acids (mainly oleic and linoleic). In terms of lipid composition, there is a greater similarity between human and rat pancreas than with other known pancreas, such as the guinea pig and the ox.

Identification by immunoblotting of somatostatin proforms in a rat pancreatic cell line.

The immunoblotting technique is applied to the analysis of "somatostatin" compounds secreted by R.I.N. T3 cells. We can confirm that two proforms of 15,300 +/- 750 and 29,000 +/- 1,100 Da accumulate in the extracellular medium. The unexpected form of 29 kDa, probably a dimeric form, disappears in reducing conditions. However, the 15 kDa peptide is characterized by several antibodies directed against either the intramolecular cycle of somatostatin-14 or the N-terminal extension of somatostatin-28. The 15 kDa form presents the same electrophoretic mobility in SDS-PAGE than the prosomatostatin isolated from a hypothalamic extract. Furthermore, this compound corresponds to the calculated mass of 10,388 Da deduced from the cDNA sequence. The detection of an immunoreactive 6 kDa peptide in the gel filtration fractions suggests an intermediate step in the prosomatostatin processing in these cells.

Studies on human pancreatic acini: action of secretagogues on amylase release and cellular cyclic AMP accumulation.

A technique for preparing a suspension of dispersed functional acini from human pancreas has been developed. The changes in pancreatic enzyme secretion and accumulation of cellular cyclic AMP caused by various secretagogues have been studied. Ca2+-mobilizing agents stimulated amylase release from human pancreatic acini. The relative potencies with which secretagogues increased amylase release were as follows: gastrin-releasing peptide's potency (Ec50, 0.1 +/- 0.01 nM) was greater than bombesin 14's (Ec50, 0.2 +/- 0.01 nM), which was greater than litorin's (Ec50, 0.6 +/- 0.18 nM), which was greater than bombesin 9's (Ec50, 6 +/- 0.1 nM). For CCK-peptides, the relative potencies were as follows: CCK-39's potency (Ec50, 0.28 +/- 0.15 microM) was equal to cerulein's (Ec50, 0.3 +/- 0.07 microM). Both of these potencies were greater than CCK-8's (Ec50, 1.6 +/- 0.1 microM), which was greater than that of CCK-4. Carbamyl choline was poorly potent (Ec50 greater than 1 mM). The 12-O-tetradecanoylphorbol-13-acetate (TPA) was active from 0.1 nM to 0.1 microM. Neither secretin nor VIP increased amylase release from human pancreatic acini but they did cause an accumulation of cellular cyclic AMP, secretin (Ec50, 0.5 +/- 0.2 nM) being more potent than VIP.

The decrease of the non secretory phospholipase A in rat pancreas during a chronic alcohol intoxication.

It is known that ethanol induces morphological lesions in the exocrine pancreas of man and of experimental animals. We showed recently that ethanol is metabolized by the rat pancreas. It has also been demonstrated that ethanol acts on the lipid metabolism of the pancreas by stimulating the lipid biosynthesis and by inhibiting fatty acids oxidation. We recently characterised a non secretory phospholipase A in the rat pancreas, probably involved in the intracellular phospholipid turnover. The actions of chronic alcoholic intoxication on the level of this enzyme is investigated in this paper. The ethanol intoxication was prolonged for two years and resulted in a progressive decrease in the level of the pancreatic non secretory phospholipase (p less than 0.01). This result confirms the chronic metabolic modifications induced by alcohol on the pancreas and emphasizes its metabolic participation in chronic alcoholic pancreatitis.

[Inhibition of the adherence of leukocytes labelled with Cr-51 in colorectal cancer]. / L'inhibition de l'adhérence des leucocytes marqués au 51Cr dans le cancer colo-rectal.

The leukocyte adherence inhibition test (LAI) is an in vitro test of immunity. Its high sensitivity in early cancer and its organ-specificity are of great interest in the study of human tumors. We developed an isotopic method (51Cr-LAI) for colorectal cancer, which has the advantage of being easier to perform than optic-counting assays. 87 patients (35 colorectal cancers, 29 benign diseases, 23 cancers of non-colorectal origin) were studied. We obtained a statistically significant difference between the mean LAI index for colorectal cancers and that for controls (p less than 0.01). LAI values greater than 13 p. 100 were considered positive. The specificity of the test was 94 p. 100 and the sensitivity for all colorectal cancers was 66 p. 100 with the highest value in the Dukes A group (LAI = 83 p. 100). Tests were negative in disseminated neoplasms. The actual fields of interest of LAI in tumor immunology and clinical research are discussed.

[Transfection of pancreatic acinar cells (AR4-2J) by bFGF modifies cell morphology and biosynthesis of pancreatic secretory enzymes]. / La transfection des cellules acineuses pancréatiques (AR4-2J) par le FGFb modifie la morphologie cellulaire et la biosynthèse des enzymes pancréatiques sécrétoires.

Basic fibroblast growth factor (bFGF or FGF-2) is present in the basal membrane of pancreatic cells during the pancreatic embryonic development. The expression of bFGF receptors has been described in normal pancreatic cells. By contrast, pancreatic cancer cells express not only the bFGF receptors but also the bFGF itself. With the aim of understanding the effects induced by the production of bFGF by pancreatic cancer cells, the pancreatic acinar cell line (AR4-2J) was used. AR4-2J cells do not produce bFGF but express bFGF receptors. These cells were transfected with a vector containing the bFGF cDNA encoding the three different forms of bFGF characterized in tumor cells. Results showed that the bFGF expression induced important phenotypic and enzymatic modifications. The transfected cells lost some morphological features of the acinar cells and expressed amylase and lipase at low levels (a 90% decrease for amylase activity, whereas lipase activity was barely detectable). These results suggest that bFGF could be involved in maintaining pancreatic cells in a slightly differentiated state.