Identification of Biomarkers for Non-small-cell Lung Cancer Patients Treated With an Immune Checkpoint Inhibitor.

BACKGROUND/AIM: Immune checkpoint inhibitors (ICIs) have an important role in lung cancer therapy. Although the programmed cell death protein-1 (PD-L1) tumor proportion score (TPS) and tumor mutational burden are known prognostic factors, they are insufficient to predict clinical outcomes. This study was conducted to identify novel biomarkers for ICI treatment. PATIENTS AND METHODS: We performed univariable and multivariable analyses of 110 patients with advanced non-small-cell lung cancer (NSCLC) who were treated with an ICI to identify novel biomarkers related to prognosis. We assessed their backgrounds, such as performance status (PS), PD-L1 TPS, smoking status, and peripheral white blood cell counts at baseline and on the day the second course of ICI administration. RESULTS: In the multivariable analysis, PS, driver gene, immune-related adverse events, and post-treatment absolute neutrophil counts (post-ANCs) were significantly associated with progression-free survival. CONCLUSION: A high level of post-ANCs was associated with poor outcome in ICI-treated NSCLC patients.

Synchronous Detection of Circulating Tumor Cells in Blood and Disseminated Tumor Cells in Bone Marrow Predicts Adverse Outcome in Early Breast Cancer.

PURPOSE: We examined the prognostic impact of circulating tumor cells (CTCs) and disseminated tumor cells (DTCs) detected at the time of surgery in 742 untreated patients with early breast cancer. EXPERIMENTAL DESIGN: DTCs in bone marrow were enumerated using the EPCAM-based immunomagnetic enrichment and flow cytometry (IE/FC) assay. CTCs in blood were enumerated either by IE/FC or CellSearch. Median follow-up was 7.1 years for distant recurrence-free survival (DRFS) and 9.1 years for breast cancer-specific survival (BCSS) and overall survival (OS). Cox regressions were used to estimate hazard ratios for DRFS, BCSS, and OS in all patients, as well as in hormone receptor-positive (HR-positive, 87%) and HR-negative (13%) subsets. RESULTS: In multivariate models, CTC positivity by IE/FC was significantly associated with reduced BCSS in both all (n = 288; P = 0.0138) and HR-positive patients (n = 249; P = 0.0454). CTC positivity by CellSearch was significantly associated with reduced DRFS in both all (n = 380; P = 0.0067) and HR-positive patients (n = 328; P = 0.0002). DTC status, by itself, was not prognostic; however, when combined with CTC status by IE/FC (n = 273), double positivity (CTC+/DTC+, 8%) was significantly associated with reduced DRFS (P = 0.0270), BCSS (P = 0.0205), and OS (P = 0.0168). In HR-positive patients, double positivity (9% of 235) was significantly associated with reduced DRFS (P = 0.0285), BCSS (P = 0.0357), and OS (P = 0.0092). CONCLUSIONS: Detection of CTCs in patients with HR-positive early breast cancer was an independent prognostic factor for DRFS (using CellSearch) and BCSS (using IE/FC). Simultaneous detection of DTCs provided additional prognostic power for outcome, including OS.

Role of C16, angiopoietin-1 and regeneration gene protein 2 in attenuating inflammation in an experimental rat model of autoimmune encephalomyelitis.

Multiple sclerosis (MS) is a chronic neurological disorder that affects the central nervous system (CNS), and results in CNS inflammation and damage to myelin. In this study, we examined the possible synergistic effects of C16, angiopoietin-1 (Ang-1) and regeneration gene protein 2 (Reg-2) in alleviating inflammation in an acute experimental autoimmune encephalomyelitis (EAE) model. We employed multiple histological, morphological and iconographic assays to examine the effect of those drugs on disease onset, clinical scores and behavioral deficits. Our results demonstrated that triple combination therapy was more efficient than the monotherapy in EAE treatment. The triple therapy significantly delayed the onset of motor symptoms, reduced disease severity, attenuated inflammatory cell infiltration and suppressed the secretion of proinflammatory cytokines. Additionally, treatment increased anti-inflammatory cytokines expression, inhibited reactive astrocytes proliferation, reduced demyelination and axonal loss, and finally reduced the neural death. Specifically, Reg-2 administration rescued oligodendrocytes and neuronal axons mainly by direct neurotrophic effects, while C16+Ang-1 (C+A) mainly improved the inflammatory milieu. In conclusion, our study suggests a possible synergistic effect through targeting a variety of pathways in relieving the clinical symptoms of inflammation in acute EAE model. Therefore, using molecules that target different molecular pathways can be beneficial for exploring novel therapeutic approaches for MS treatment.

Genomic profiling for copy number changes in plasma of ovarian cancer patients - a new era for cancer diagnostics?

A blood test that can detect human malignancy with high clinical sensitivity and specificity is highly desirable. To achieve this, a tumor marker is needed that correlates with tumor burden and that can be measured with high analytical sensitivity and specificity. Over the past decades, a number of different types of tumor markers have emerged, including proteins such as enzymes, glycoproteins, and oncofetal antigens. Besides proteins, genetic abnormalities such as mutations, amplifications, and circulating tumor DNA have served as tumor markers. Despite the diversity of such biomarkers, their acceptance and implementation into routine clinical practice requires that their use results in improvements in patient outcome. Current tumor markers used in the clinic have limited utility. As such, innovative approaches to identifying tumor markers are highly desirable and one such approach may be to look for sub-chromosomal changes in the blood of patients with ovarian cancer, as is routinely performed in prenatal screening.Please see related article: http://bmcmedicine.biomedcentral.com/articles/10.1186/s12916-016-0667-6.

(68)Ga-DOTA-peptide: A novel molecular biomarker for nasopharyngeal carcinoma.

BACKGROUND: Increased somatostatin receptor (SSTR) expression in patients with undifferentiated nasopharyngeal carcinoma (NPC) has been demonstrated with receptor autoradiography, (111) In-Octreotide scintigraphy, and (68) Ga-DOTA-TOC positron emission tomography (PET)/CT imaging. We sought to compare and correlate the uptake of fluorodeoxyglucose (FDG) and DOTA-NOC in undifferentiated NPC to ascertain the possible role of (68) Ga-DOTA-NOC PET/CT as a new imaging biomarker and to assess whether targeted peptide receptor radionuclide therapy is a feasible treatment option. METHODS: After obtaining approval from our institutional review board, 4 patients with biopsy proven nonkeratinizing undifferentiated NPC who had just undergone routine staging/restaging (18) F-FDG PET/CT imaging were prospectively and consecutively recruited for (68) Ga-DOTA-NOC PET/CT imaging. Of these 4 patients, 3 were newly diagnosed with untreated NPC, whereas 1 patient was diagnosed with a case of recurrent NPC with previous treatment. These patients subsequently underwent (68) Ga-DOTA-NOC PET/CT within 10 days from the (18) F-FDG PET/CT to ensure lesion comparability. Tracer uptake in tumor lesions were assessed visually and semiquantitatively by measuring maximum standardized uptake values (SUVmax). RESULTS: There were 12 FDG-avid lesions of which 7 showed avid uptake of DOTA-NOC greater than liver uptake, whereas 5 showed low uptake of DOTA-NOC less than liver uptake. Subset analysis of the FDG-avid lesions at the primary and recurrent sites showed that all the FDG-avid primary tumors in the nasopharynx showed avid uptake of DOTA-NOC. On the contrary, the case of recurrent NPC showed avid FDG uptake but low DOTA-NOC uptake. Subset analysis of the suspicious FDG-avid cervical lymph nodes showed that 50% of them demonstrated avid DOTA-NOC uptake greater than liver uptake, whereas the remaining demonstrated low-grade DOTA-NOC uptake less than liver uptake. The 2 subcentimeter cervical lymph nodes that showed low-grade uptake of FDG lower than mediastinal blood pool activity were deemed to be reactive/inflammatory and showed low-grade uptake of DOTA-NOC. CONCLUSION: This study highlights the potential of (68) Ga-DOTA-peptide PET/CT as a new molecular biomarker for newly diagnosed undifferentiated NPC, and less so for recurrent NPC and metastatic nodes. This potentially opens up new diagnostic and therapeutic options in the management of undifferentiated NPC.

Recombinant Reg3α protein protects against experimental acute pancreatitis in mice.

Regenerating gene 3α (Reg3α) protein is a trophic factor that stimulates cell and tissue proliferation, neogenesis and also acts against apoptosis and necrosis. In order to explore the potential roles of recombinant Reg3α (rReg3α), we produced a mature rReg3α polypeptide for direct administration in l-arginine (L-Arg) induced acute pancreatitis (AP) in mice. Our results showed that rReg3α stimulated cell proliferation through Erk1/2 and p38 phosphorylation and also cyclin D1 upregulation mediated by Akt/ATF-2 signaling. Moreover, rReg3α administration significantly reduced the pancreatic damage caused by L-Arg injection, as shown in histological examination and serum amylase, lipase and C-reactive protein (CRP) assays. Not only acinar cell necrosis but also apoptosis found in the pancreas of AP mice were alleviated by rReg3α. Finally, upregulated Bcl-2 and Bcl-xL and suppressed poly (ADP-ribose) synthetase/polymerase (PARP) levels were detected as being relevant to the mechanism of rReg3α protection. We therefore conclude that rReg3α acts as a protective polypeptide against AP in mice by enhancing Bcl-2 and Bcl-xL expressions and suppressing PARP level.

Effect of novel chitosan-fluoroaluminosilicate resin modified glass ionomer cement supplemented with translationally controlled tumor protein on pulp cells.

Dental materials that can promote cell proliferation and function is required for regenerative pulp therapy. Resin modified glass ionomer cement (RMGIC), a broadly used liner or restorative material, can cause apoptosis to pulp cells mainly due to HEMA (2-hydroxyethyl methacrylate), the released residual monomer. Recent studies found that chitosan and albumin could promote release of protein in GIC while translationally controlled tumor protein (TCTP) has an anti-apoptotic activity against HEMA. The aim of this study was to examine the effect of chitosan and albumin modified RMGIC (Exp-RMGIC) supplemented with TCTP on pulp cell viability and mineralization. Exp-RMGIC+TCTP was composed of RMGIC powder incorporated with 15 % of chitosan, 5 % albumin and supplemented with TCTP mixed with the same liquid components of RMGIC. The effect of each specimen on pulp cells was examined using the Transwell plate. From the MTT assay, Exp-RMGIC+TCTP had the highest percentages of viable cells (P < 0.05) at both 24 and 74 h. Flow cytometry revealed that, after 24 h, Exp-RMGIC+TCTP gave the lowest percentages of apoptotic cells compared to other groups. There was no difference in alkaline phosphatase (ALP) activity among different formula of the specimens, while cells cultured in media with TCTP had higher ALP activity. Von Kossa staining revealed that RMGIC+TCTP, and Exp-RMGIC+TCTP had higher percentages of calcium deposit area compared to those without TCTP. It was concluded that Exp-RMGIC supplemented with TCTP had less cytotoxicity than RMGIC and can protect cells from apoptosis better than RMGIC supplemented with TCTP.

Análise morfológica da mucosa oral de ratos submetida a carcinogênese experimental pelo àxido de nitroquinolina (4NQO) / Morphological analysis of the oral mucosa of rats subjected to experimental carcinogenesis by administration of nitroquinoline oxide (4NQO)

Objetivo: Induzir a carcinogênese na mucosa lingual de ratos Wistar através do óxido de nitroquinolina (4NQO), relacionando as alterações clínicas e microscópicas desenvolvidas com o tempo de exposição ao carcinógeno. Método: Foram utilizados 20 ratos que foram distribuídos entre o grupo experimental (15 animais) e controle (5 animais). No grupo experimental os animais foram alocados em 3 subgrupos, nos quais 5 animais receberam o 4NQO por 2 meses (A1), 5 por 3 meses (A2) e o restante por 4 meses (A3), o que tornou possível a detecção das diferentes fases da carcinogênese. Os dados foram apresentados por meio da estatística descritiva e foram utilizados os testes de Kruskal-Wallis e de Mann-Whitney. O programa estatístico utilizado foi o SPSS (Statistical Package for the Social Sciences) na versão 13 e a margem de erro utilizada foi de 5,0%. Resultados: Tanto o modelo animal escolhido, como o carcinógeno químico utilizado foram satisfatórios para produção da carcinogênese oral similar a que ocorre em humanos. As alterações clínicas foram mais acentuadas nos animais que receberam as aplicações tópicas do 4NQO durante 4 meses, sendo possível visualizar desde uma leucoplasia a regiões eritroleucoplásicas. As alterações histopatológicas observadas no epitélio oral foram compatíveis com o diagnóstico de displasia epitelial leve, moderada, severa, carcinoma in situ ou carcinoma invasivo. Conclusão: A severidade das lesões teve relação direta com o tempo de exposição ao 4NQO.

Objective: To induce carcinogenesis on the lingual mucosa of Wistar rats by the administration of nitroquinoline oxide (4NQO), relating the clinical and microscopic alterations developed during the time of exposure to the carcinogenic agent. Methods: Twenty rats were allocated to either an experimental (15 animals) or a control (5 animals) group. In the experimental group, the animals were divided into 3 subgroups in which 5 animals received 4NQO during 2 months (A1), 5 during 3 months (A2), and 5 during 4 months (A3), making it possible to detect the different phases of carcinogenesis. Data were presented as descriptive statistics using the Kruskal-Wallis and Mann-Whitney tests in the SPSS (Statistical Package for the Social Sciences) software, version 13.0. The significance level was set at 5.0%. Results: Both the animal model and the carcinogenic agent used in the study were satisfactory for inducing similar oral carcinogenesis to the one occurring in humans. The clinical alterations were more accentuated in the animals that received the topical applications of 4NQO during 4 months, ranging from leukoplakia to erythroleukoplakias regions. The histopathological alterations observed in the oral epithelium were compatible with the diagnosis of mild, moderate and severe epithelial dysplasia, in situ carcinoma or invasive carcinoma. Conclusion: The severity of the lesions had a direct relationship with the exposure time to 4NQO.

Análise Morfológica da Mucosa Oral de Ratos Submetida à Carcinogênese Experimental pelo Óxido de Nitroquinolina (4NQO) / Morphological Analysis of the Oral Mucosa of Rats Subjected to Experimental Carcinogenesis by Administration of Nitroquinoline Oxide (4NQO)

Objetivo: Induzir a carcinogênese na mucosa lingual de ratos Wistar através do óxido de nitroquinolina (4NQO), relacionando as alterações clínicas e microscópicas desenvolvidas com o tempo de exposição ao carcinógeno. Método: Foram utilizados 20 ratos que foram distribuídos entre o grupo experimental (15 animais) e controle (5 animais). No grupo experimental os animais foram alocados em 3 subgrupos, nos quais 5 animais receberam o 4NQO por 2 meses (A1), 5 por 3 meses (A2) e o restante por 4 meses (A3), o que tornou possível a detecção das diferentes fases da carcinogênese. Os dados foram apresentados por meio da estatística descritiva e foram utilizados os testes de Kruskal-Wallis e de Mann-Whitney. O programa estatístico utilizado foi o SPSS (Statistical Package for the Social Sciences) na versão 13 e a margem de erro utilizada foi de 5,0%. Resultados: Tanto o modelo animal escolhido, como o carcinógeno químico utilizado foram satisfatórios para produção da carcinogênese oral similar a que ocorre em humanos. As alterações clínicas foram mais acentuadas nos animais que receberam as aplicações tópicas do 4NQO durante 4 meses, sendo possível visualizar desde uma leucoplasia a regiões eritroleucoplásicas. As alterações histopatológicas observadas no epitélio oral foram compatíveis com o diagnóstico de displasia epitelial leve, moderada, severa, carcinoma in situ ou carcinoma invasivo. Conclusão: A severidade das lesões teve relação direta com o tempo de exposição ao 4NQO.

Objective: To induce carcinogenesis on the lingual mucosa of Wistar rats by the administration of nitroquinoline oxide (4NQO), relating the clinical and microscopic alterations developed during the time of exposure to the carcinogenic agent. Methods: Twenty rats were allocated to either an experimental (15 animals) or a control (5 animals) group. In the experimental group, the animals were divided into 3 subgroups in which 5 animals received 4NQO during 2 months (A1), 5 during 3 months (A2), and 5 during 4 months (A3), making it possible to detect the different phases of carcinogenesis. Data were presented as descriptive statistics using the Kruskal-Wallis and Mann-Whitney tests in the SPSS (Statistical Package for the Social Sciences) software, version 13.0. The significance level was set at 5.0%. Results: Both the animal model and the carcinogenic agent used in the study were satisfactory for inducing similar oral carcinogenesis to the one occurring in humans. The clinical alterations were more accentuated in the animals that received the topical applications of 4NQO during 4 months, ranging from leukoplakia to erythroleukoplakias regions. The histopathological alterations observed in the oral epithelium were compatible with the diagnosis of mild, moderate and severe epithelial dysplasia, in situ carcinoma or invasive carcinoma. Conclusion: The severity of the lesions had a direct relationship with the exposure time to 4NQO.

The neuroprotective effects of Reg-2 following spinal cord transection injury.

This study was designed to elucidate the potential neuroprotective effects of Reg-2 (regeneration gene protein 2) in a rodent model of spinal cord transection injury at the ninth thoracic level. Reg-2 at 100 and 500 µg, recombinant rat ciliary neurotrophic factor, or vehicle were delivered intrathecally using Alzet miniosmotic pumps. We found that Reg-2 treatment significantly reduced neuronal death in the spinal cord. There was also an attenuation of inflammation at the injury site and an increase in white matter sparing and retained myelination. Retrograde tracing revealed that Reg-2 protected axons of long descending pathways at 6 weeks post-SCI, and the number of FluoroGold-labeled neurons in spinal and supraspinal regions was also significantly increased. Immunofluorescent staining confirmed that the spared white matter contained neurofilament-positive axons. Moreover, behavioral improvements were revealed by Basso Beattie Bresnahan locomotor rating scores and grid-walk analysis. These results suggest that Reg-2 might promote functional recovery by increasing axonal growth, inhibiting neuronal apoptosis, and attenuating spinal cord secondary injury after SCI.

MutS homologue 2 and the long-term benefit of adjuvant chemotherapy in lung cancer.

PURPOSE: We sought to determine the long-term (median follow-up, 7.5 years) predictive power of human MutS homologue 2 (MSH2) immunohistochemical expression in patients who enrolled in the International Adjuvant Lung Trial. EXPERIMENTAL DESIGN: We tested the interaction between MSH2 and the allocated treatment (chemotherapy versus observation) in a Cox model adjusted on clinicopathologic variables. The significance level was set at 0.01. RESULTS: MSH2 levels were low in 257 (38%) and high in 416 (62%) tumors. The benefit from chemotherapy was likely different according to MSH2 (interaction test, P = 0.06): there was a trend for chemotherapy to prolong overall survival when MSH2 was low [hazard ratio (HR), 0.76; 95% confidence interval (95% CI), 0.59-0.97; P = 0.03], but not when MSH2 was high (HR, 1.12; 95% CI, 0.81-1.55; P = 0.48). In the control arm, the HR was 0.66 (95% CI, 0.49-0.90; P = 0.01) when MSH2 was high. When combining MSH2 with excision repair cross-complementing group 1 (ERCC1) into four subgroups, the benefit of chemotherapy decreased with the number of markers expressed at high levels (P = 0.01). A similar decrease was noted when combining MSH2 and P27 (P = 0.01). Chemotherapy prolonged overall survival in the combined low MSH2/low ERCC1 subgroup (HR, 0.65; 95% CI, 0.47-0.91; P = 0.01) and in the combined low MSH2/low P27 subgroup (HR, 0.65; 95% CI, 0.46-0.93; P = 0.01). CONCLUSIONS: MSH2 expression is a borderline significant predictor of a long-term benefit from adjuvant cisplatin-based chemotherapy in patients with completely resected lung cancer. MSH2 combined with ERCC1 or P27 may identify patients most likely to benefit durably from chemotherapy.

Islet neogenesis-associated protein pentadecapeptide (INGAP-PP): mechanisms involved in its effect upon beta-cell mass and function.

The effect of islet neogenesis-associated protein pentadecapeptide (INGAP-PP) administration to normal male hamsters upon serum glucose and triglyceride levels, beta-cell mass and function was studied. INGAP-PP (500 mug) or saline was injected twice daily during 10 days. Both groups showed comparable body weight, serum glucose and triglyceride levels. INGAP-PP treated animals had significantly higher HOMA-IR and HOMA-beta and their islets released more insulin in response to glucose; they had lower islet DNA content, significantly increased number of islets/unit area, beta-cell replication rate and mass, cells co-expressing Pdx-1/INGAP and islets in contact with ducts, and decreased beta-cell apoptosis rate. The percentage of cells expressing Pdx-1 alone or together with INGAP or insulin increased significantly in ducts. These animals also showed a significantly higher concentration of Pdx-1 and Ngn-3 mRNA and a lower number of INGAP-positive cells. In conclusion, INGAP-PP promoted a controlled and functionally active increase of beta-cell mass; our data demonstrate for the first time the mechanism responsible for such changes; that Ngn-3 would be involved in INGAP-PP-induced neogenesis; and the existence of a negative feedback loop with endogenous INGAP-producing cells. Accordingly, INGAP-PP could be used to induce these effects in people with or at risk of developing diabetes.

Composite hydrogel formulations of stratifin to control MMP-1 expression in dermal fibroblasts.

PURPOSE: Stratifin is a potent anti-fibrogenic factor that stimulates the expression of matrix metalloproteinase-1 (MMP-1) in dermal fibroblasts. The propose of this work was to develop a controlled release delivery system for stratifin that can be applied at the time of wound closure to release stratifin and stimulate the expression of MMP-1 in a sustained manner over the late stages of wound healing (after 3 days). METHODS: Stratifin was complexed to chitosan particles, which were then encapsulated in PLGA microspheres and blended into crosslinked hyaluronic acid films. In vitro release was assessed using fluorescent-tagged stratifin, cytotoxicity by MTT assay and bioactivity by measuring the levels of MMP-1 expression in cultured fibroblasts. RESULTS: The release of stratifin was delayed for 3 days and then controlled for 30 days so that 60% of the total stratifin loaded was released. The released protein significantly stimulated the expression of MMP-1 in cultured fibroblasts without compromising cell viability. By complexing to chitosan, the initial burst release was reduced, so that only 5% of stratifin was released in 3 days. CONCLUSION: This stratifin delivery system has the potential to be used as an anti-fibrogenic factor-associated wound insert for improving post-surgical scarring in closed wound.

Marginal zone variant of mantle cell lymphoma: CD5-negative cyclin D1-positive variant posing a diagnostic dilemma.

Described herein is an unusual case of mantle cell lymphoma (MCL) histologically mimicking marginal zone lymphoma (MZL). An 83-year-old man presented with multiple adenopathies and a hilar mass encroaching on the right lung. A transbronchial biopsy showed small blue cells suspicious for small cell carcinoma. On further analysis the cells were predominantly small cleaved and CD20 positive, suggesting follicular lymphoma, grade 2. An axillary lymph node biopsy showed germinal centers surrounded by monocytoid B cells. Flow cytometry was negative for CD5 and CD23 and the diagnosis of MZL was considered. Because of the aggressive clinical behavior, including extensive necrosis on imaging studies, immunohistochemistry for cyclin D-1 was performed and was positive. Bone marrow was extensively involved and it showed t(11;14), in addition to other complex cytogenetic abnormalities. Differentiating MCL from MZL has prognostic and therapeutic implications, particularly when considering the potential role of targeted therapy and cell cycle modulators.

[Effect of 14-3-3sigma on transcriptional activity of p73 gene].

BACKGROUND & OBJECTIVE: p53 gene is the main regulator of 14-3-3sigma. Activated p53 could induce the expression of 14-3-3sigma, while 14-3-3sigma stabilizes the expression of p53 and enhances its transcriptional activity. p63 and p73, the members of p53 family, also have some functions similar to p53. This study was to investigate the effect of 14-3-3sigma on the transcriptional activity of p73. METHODS: Luciferase reporter assay, reverse transcription-polymerase chain reaction (RT-PCR), and Western blot were used to evaluate the effect of 14-3-3sigma on the transcriptional activity of p73 in p53-deficient human lung carcinoma cell line H1299. Colony formation test was used to evaluate the effect of 14-3-3sigma on the transcriptional activity of p73 in p53-mutant human breast cancer cell line MDA-MB-436. RESULTS: The luciferase activities induced by bax and p21WAF1 promotors were significantly higher in p73-transfected H1299 cells than in control H1299 cells (P<0.01), and were further increased by the transfection of p73 (25 ng) and 14-3-3sigma (100, 200, and 400 ng) in a dose-dependent manner (P<0.01). The expression of bax and p21WAF1 were higher in p73-transfected H1299 cells than in control H1299 cells, and were significantly higher in p73-and 14-3-3sigma-transfected H1299 cells than in p73-transfected H1299 cells (P<0.01). The number of colonies was fewer in p73-transfected MDA-MB-436 cells than in control MDA-MB-436 cells, and the colonies were significantly smaller in p73-and 14-3-3sigma-transfected H1299 cells than in p73-transfected H1299 cells (P<0.01). CONCLUSION: 14-3-3sigma can enhance the transcriptional activity of p73 in a dose-dependent manner.

Calbindin-d(28k): a marker of recurrence for medulloblastomas.

BACKGROUND: The expression of the Ca(2+)-binding protein calbindin-D(28k) was analyzed in medulloblastomas in relation to clinical features and other biologic markers related to cell proliferation, differentiation, p53, and cerebellar developmental regulated gene expression. METHODS: Immunohistochemistry was carried out on histologic slides from a first retrospective series of 29 nonmetastatic and 10 metastatic medulloblastoma formalin-fixed, paraffin-embedded tissues, using specific antibodies against calbindin-D(28k), calretinin, alpha-parvalbumin and beta-parvalbumin, and S100 proteins. Informed consent was obtained from the subjects and/or guardians. Other biologic markers for differentiation, cell proliferation, the expression of the p53 tumor suppressor gene protein, and cerebellar developmental regulated genes were similarly investigated. A second series of 16 medulloblastomas from young patients (younger than 15 years) was added in order to validate the results obtained in the first series. RESULTS: Of all the markers investigated, only calbindin-D(28k) was significantly associated with prognosis. Survival and remission (i.e. recurrence free) time analysis performed on all the cases (n = 55) confirmed a high risk of death (P = 0.004) and recurrence (P = 0.003) associated with calbindin-positivity. As calbindin-positivity was predominantly observed in tumors from young patients, the authors confirmed its prognostic value in the subgroup of patients younger than 15 years (n = 37). Cox regression analysis showed a significant and independent prognostic value for calbindin expression and, to a lesser extent, the type of surgery (total or subtotal). Three risk groups were thus identified, distinguishing among the cases characterized by a total resection and calbindin-negativity (good prognosis), by a subtotal resection and calbindin-negativity (intermediary), and by calbindin-positivity (bad prognosis). CONCLUSIONS: The current study suggests that calbindin-positive medulloblastomas represent a subclass of aggressive tumors more frequently seen in younger patients.

[Improvement of prescriptions for serum tumor markers in a general hospital]. / Amélioration de la prescription de marqueurs tumoraux sériques dans un hôpital général.

OBJECTIVES: Using a standardized prescription sheet we attempted to improve requests for serum tumor markers in a general hospital. METHODS: Over two 35-day periods before and one year after defining a local prescription consensus and introducing a new prescription sheet, we counted the number of orders for five tumor markers (CEA, CA 19-9, CA 15-3, CA 125, alpha FP) and determined their compliance to the defined prescription protocol. RESULTS: Between the two study periods, the number of prescriptions for the designated tumor markers fell by 24%, from 153 requests in 94 patients to 123 requests in 99 patients, despite a 6% increase in the number of admissions. There was a significant reduction in the number of serum markers orders per prescription (from 1.6 to 1.2) although the distribution by tumor marker remained unchanged. Compliance to the prescription protocol improved, rising from 65 to 87% in units where the pre-protocol compliance rate was below 80%. The rate of compliance was not correlated with correct completion of the new prescription sheet (91% vs 86% respectively). The 6-month cost-savings was estimated at 31,104 FF using the general French nomenclature for laboratory tests. Direct cost reduction was estimated at 5,688 FF. CONCLUSION: Long-lasting improvement of serum tumor marker prescriptions can be achieved in a general hospital. Obtaining a local consensus implicating all prescribing units seems more important than a change in the presentation of the prescription sheet.