Enhanced antitumor effect of L-buthionine sulfoximine or ionizing radiation by copper complexes with 2,2´-biquinoline and sulfonamides on A549 2D and 3D lung cancer cell models.

Two ternary copper(II) complexes with 2,2'-biquinoline (BQ) and with sulfonamides: sulfamethazine (SMT) or sulfaquinoxaline (SDQ) whose formulae are Cu(SMT)(BQ)Cl and Cu(SDQ)(BQ)Cl·CH3OH, in what follows SMTCu and SDQCu, respectively, induced oxidative stress by increasing ROS level from 1.0 µM and the reduction potential of the couple GSSG/GSH2. The co-treatment with L-buthionine sulfoximine (BSO), which inhibits the production of GSH, enhanced the effect of copper complexes on tumor cell viability and on oxidative damage. Both complexes generated DNA strand breaks given by-at least partially-the oxidation of pyrimidine bases, which caused the arrest of the cell cycle in the G2/M phase. These phenomena triggered processes of apoptosis proven by activation of caspase 3 and externalization of phosphatidylserine and loss of cell integrity from 1.0 µM. The combination with BSO induced a marked increase in the apoptotic population. On the other hand, an improved cell proliferation effect was observed when combining SDQCu with a radiation dose of 2 Gy from 1.0 µM or with 6 Gy from 1.5 µM. Finally, studies in multicellular spheroids demonstrated that even though copper(II) complexes did not inhibit cell invasion in collagen gels up to 48 h of treatment at the higher concentrations, multicellular resistance outperformed several drugs currently used in cancer treatment. Overall, our results reveal an antitumor effect of both complexes in monolayer and multicellular spheroids and an improvement with the addition of BSO. However, only SDQCu was the best adjuvant of ionizing radiation treatment.

Cytotoxicity and DNA damage evaluation of TiO2 and ZnO nanoparticles. Uptake in lung cells in culture.

The cytotoxicity and DNA damage of titanium dioxide and zinc oxide nanoparticles (TiO2 and ZnO NPs) have been studied in a human lung carcinoma cell line (A549) after 24 h exposure. TiO2 and ZnO NPs had mean diameters of 12.9 ± 2.8 and 24.1 ± 8.0 nm, respectively. ZnO NPs reduced cell viability from 250 µg/mL, increasing reactive oxygen species (ROS) and decreased GSH/GSSG ratio. The comet assay detected DNA damage from 50 µg/mL. TiO2 NPs induced cytotoxicity and DNA damage from 50 to 100 µg/mL, respectively, along with a decrease of the GSH/GSSG ratio. Both particles were found inside the cells, within membrane-bound vesicles. The internalization mechanism is promoted partially by caveolae-mediated endocytosis and, in the case of TiO2 NPs, also by macropinocytosis.

Síndrome de CLOVES: reporte de un caso y revisión de los diagnósticos diferenciales / CLOVES syndrome: case report and review of differential diagnoses

Resumen Presentamos el caso de una paciente de 27 años con diagnóstico reciente de síndrome de CLOVES (Congenital, Lipomatous, Overgrowth, Vascular malformations, Epidermalnevi and Spinal/Skeletalanomalies and/orScoliosisSyndrome), quien fue diagnosticada previamente con los síndromes Klippel-Trenaunay-Weber y de Proteus. El síndrome de CLOVES es una patología poco frecuente y muchas veces el diagnóstico basado en la clínica suele complicarse por la superposición de signos y síntomas con otras patologías que también cursan con sobrecrecimiento.

Abstract We present the case of a 27-years-old patient with a newly diagnosis of CLOVES syndrome (Congenital, Lipomatous, Overgrowth, Vascular malformations, Epidermal nevi and Spinal/Skeletal anomalies and/or Scoliosis Syndrome). She has previously been diagnosed of Klippel-Trenaunay-Weber (at birth) and Proteus Syndrome (at 7 years). She presents dermatological alterations, syndactyly and overgrowth. CLOVES syndrome is a rare disease and often the clinic-based diagnostic is difficult due to overlapping signs and symptoms with other illnesses that also involve overgrowth.

6-Methoxyquinoline complexes as lung carcinoma agents: induction of oxidative damage on A549 monolayer and multicellular spheroid model.

The aim of this work was to study the antitumor effects and the mechanisms of toxic action of a series of 6-methoxyquinoline (6MQ) complexes in vitro. The Cu(II) and Zn(II) complexes (Cu6MQ and Zn6MQ) are formulated as M(6MQ)2Cl2; the Co(II) and Ag(I) compounds (Co6MQ and Ag6MQ) are ionic with formulae [Ag(6MQ)2]+NO3- and H(6MQ)+[Co(6MQ)Cl3]- (where H(6MQ)+ is the protonated ligand). We found that the copper complex, outperformed the Co(II), Zn(II) and Ag(I) complexes with a lower IC50 (57.9 µM) in A549 cells exposed for 24 h. Cu6MQ decreased cell proliferation and induced oxidative stress detected with H2DCFDA at 40 µM, which reduces GSH/GSSG ratio. This redox imbalance induced oxidative DNA damage revealed by the Micronucleus test and the Comet assay, which turned into a cell cycle arrest at G2/M phase and induced apoptosis. In multicellular spheroids, the IC50 values tripled the monolayer model (187.3 µM for 24 h). At this concentration, the proportion of live/dead cells diminished, and the spheroids could not proliferate or invade. Although Zn6MQ also decreased GSH/GSSG ratio from 200 µM and the cytotoxicity is related to oxidative stress, the induction of the hydrogen peroxide levels only doubled the control value. Zn6MQ induced S phase arrest, which relates with the increased micronucleus frequency and with the induction of necrosis. Finally, our results reveal a synergistic activity with a 1:1 ratio of both complexes in the monolayer and multicellular spheroids.

Detection of generalized synchronization using echo state networks.

Generalized synchronization between coupled dynamical systems is a phenomenon of relevance in applications that range from secure communications to physiological modelling. Here, we test the capabilities of reservoir computing and, in particular, echo state networks for the detection of generalized synchronization. A nonlinear dynamical system consisting of two coupled Rössler chaotic attractors is used to generate temporal series consisting of time-locked generalized synchronized sequences interleaved with unsynchronized ones. Correctly tuned, echo state networks are able to efficiently discriminate between unsynchronized and synchronized sequences even in the presence of relatively high levels of noise. Compared to other state-of-the-art techniques of synchronization detection, the online capabilities of the proposed Echo State Network based methodology make it a promising choice for real-time applications aiming to monitor dynamical synchronization changes in continuous signals.

Nottingham prognostic index plus (NPI+) predicts risk of distant metastases in primary breast cancer.

The Nottingham prognostic index plus (NPI+) is based on the assessment of biological class combined with established clinicopathologic prognostic variables providing improved patient outcome stratification for breast cancer superior to the traditional NPI. This study aimed to determine prognostic capability of the NPI+ in predicting risk of development of distant disease. A well-characterised series of 1073 primary early-stage BC cases treated in Nottingham and 251 cases from Budapest were immunohistochemically assessed for cytokeratin (Ck)5/6, Ck18, EGFR, oestrogen receptor (ER), progesterone receptor, HER2, HER3, HER4, Mucin 1 and p53 expression. NPI+ biological class and prognostic scores were assigned using individual algorithms for each biological class incorporating clinicopathologic parameters and investigated in terms of prediction of distant metastases-free survival (MFS). The NPI+ identified distinct prognostic groups (PG) within each molecular class which were predictive of MFS providing improved patient outcome stratification superior to the traditional NPI. NPI+ PGs, between series, were comparable in predicting patient outcome between series in luminal A, basal p53 altered and HER2+/ER+ (p > 0.01) tumours. The low-risk groups were similarly validated in luminal B, luminal N, basal p53 normal tumours (p > 0.01). Due to small patient numbers the remaining PGs could not be validated. NPI+ was additionally able to predict a higher risk of metastases at certain distant sites. This study may indicate the NPI+ as a useful tool in predicting the risk of metastases. The NPI+ provides accurate risk stratification allowing improved individualised clinical decision making for breast cancer.

Markers of progression in early-stage invasive breast cancer: a predictive immunohistochemical panel algorithm for distant recurrence risk stratification.

Accurate distant metastasis (DM) prediction is critical for risk stratification and effective treatment decisions in breast cancer (BC). Many prognostic markers/models based on tissue marker studies are continually emerging using conventional statistical approaches analysing complex/dimensional data association with DM/poor prognosis. However, few of them have fulfilled satisfactory evidences for clinical application. This study aimed at building DM risk assessment algorithm for BC patients. A well-characterised series of early invasive primary operable BC (n = 1902), with immunohistochemical expression of a panel of biomarkers (n = 31) formed the material of this study. Decision tree algorithm was computed using WEKA software, utilising quantitative biomarkers' expression and the absence/presence of distant metastases. Fifteen biomarkers were significantly associated with DM, with six temporal subgroups characterised based on time to development of DM ranging from <1 to >15 years of follow-up. Of these 15 biomarkers, 10 had a significant expression pattern where Ki67LI, HER2, p53, N-cadherin, P-cadherin, PIK3CA and TOMM34 showed significantly higher expressions with earlier development of DM. In contrast, higher expressions of ER, PR and BCL2 were associated with delayed occurrence of DM. DM prediction algorithm was built utilising cases informative for the 15 significant markers. Four risk groups of patients were characterised. Three markers p53, HER2 and BCL2 predicted the probability of DM, based on software-generated cut-offs, with a precision rate of 81.1 % for positive predictive value and 77.3 %, for the negative predictive value. This algorithm reiterates the reported prognostic values of these three markers and underscores their central biological role in BC progression. Further independent validation of this pruned panel of biomarkers is therefore warranted.

Nottingham Prognostic Index Plus (NPI+): a modern clinical decision making tool in breast cancer.

BACKGROUND: Current management of breast cancer (BC) relies on risk stratification based on well-defined clinicopathologic factors. Global gene expression profiling studies have demonstrated that BC comprises distinct molecular classes with clinical relevance. In this study, we hypothesised that molecular features of BC are a key driver of tumour behaviour and when coupled with a novel and bespoke application of established clinicopathologic prognostic variables can predict both clinical outcome and relevant therapeutic options more accurately than existing methods. METHODS: In the current study, a comprehensive panel of biomarkers with relevance to BC was applied to a large and well-characterised series of BC, using immunohistochemistry and different multivariate clustering techniques, to identify the key molecular classes. Subsequently, each class was further stratified using a set of well-defined prognostic clinicopathologic variables. These variables were combined in formulae to prognostically stratify different molecular classes, collectively known as the Nottingham Prognostic Index Plus (NPI+). The NPI+ was then used to predict outcome in the different molecular classes. RESULTS: Seven core molecular classes were identified using a selective panel of 10 biomarkers. Incorporation of clinicopathologic variables in a second-stage analysis resulted in identification of distinct prognostic groups within each molecular class (NPI+). Outcome analysis showed that using the bespoke NPI formulae for each biological BC class provides improved patient outcome stratification superior to the traditional NPI. CONCLUSION: This study provides proof-of-principle evidence for the use of NPI+ in supporting improved individualised clinical decision making.

Identification of key clinical phenotypes of breast cancer using a reduced panel of protein biomarkers.

BACKGROUND: Breast cancer is a heterogeneous disease characterised by complex molecular alterations underlying the varied behaviour and response to therapy. However, translation of cancer genetic profiling for use in routine clinical practice remains elusive or prohibitively expensive. As an alternative, immunohistochemical analysis applied to routinely processed tissue samples could be used to identify distinct biological classes of breast cancer. METHODS: In this study, 1073 archival breast tumours previously assessed for 25 key breast cancer biomarkers using immunohistochemistry and classified using clustering algorithms were further refined using naïve Bayes classification performance. Criteria for class membership were defined using the expression of a reduced panel of 10 proteins able to identify key molecular classes. We examined the association between these breast cancer classes with clinicopathological factors and patient outcome. RESULTS: We confirm patient classification similar to established genotypic biological classes of breast cancer in addition to novel sub-divisions of luminal and basal tumours. Correlations between classes and clinicopathological parameters were in line with expectations and showed highly significant association with patient outcome. Furthermore, our novel biological class stratification provides additional prognostic information to the Nottingham Prognostic Index. CONCLUSION: This study confirms that distinct molecular phenotypes of breast cancer can be identified using robust and routinely available techniques and both the luminal and basal breast cancer phenotypes are heterogeneous and contain distinct subgroups.

Biology of primary breast cancer in older women treated by surgery: with correlation with long-term clinical outcome and comparison with their younger counterparts.

BACKGROUND: As age advances breast cancer appears to change its biological characteristics, however, very limited data are available to define the precise differences between older and younger patients. METHODS: Over 36 years (1973-2009), 1758 older (≥70 years) women with early operable primary breast cancer were managed in a dedicated clinic. In all, 813 underwent primary surgery and 575 good quality tumour samples were available for biological analysis. The pattern of biomarkers was analysed using indirect immunohistochemistry on tissue microarrays. Comparison was made with a previously characterised series of younger (<70 years) patients. RESULTS: There was high expression of oestrogen receptor (ER), PgR, Bcl2, Muc1, BRCA1 and 2, E-cadherin, luminal cytokeratins, HER3, HER4, MDM2 and 4 and low expression of human epidermal growth factor receptor (HER)-2, Ki67, p53, EGFR and CK17. Oestrogen receptor and axillary stage appeared as independent prognostic factors. Unsupervised partitional clustering showed six biological clusters in older patients, five of which were common in the younger patients, whereas the low ER luminal cluster was distinct in the older series. The luminal phenotype showed better breast cancer-specific survival, whereas basal and HER2-overexpressing tumours were associated with poor outcome. CONCLUSION: Early operable primary breast cancer in older women appears as a distinct biological entity, with existence of a novel cluster. Overall older women showed less aggressive tumour biology and ER appeared as an independent prognostic factor alongside the time-dependent axillary stage. These biological characteristics may explain the differences in clinical outcome and should be considered in making therapeutic decisions.

Thermal behavior, powder X-ray diffraction, DFT calculations and vibrational spectra of barium bis(pentacyanonitrosylchromate) octahydrate, Ba3[Cr(CN)5NO]2.8H2O(D2O).

The cell parameters of Ba3[Cr(CN)5NO]2.8H2O were determined from powder X-ray diffraction using the autoindexing program TREOR, and refined by the Le Bail methods with the FULLPROF program. An orthorhombic cell was determined with cell parameters a = 15.0324(2) A, b = 12.9542(9) A, and c = 7.5094(5) A. Two possible space groups are consistent with the systematic absences: Pmcb (#55) and P2cb (#32). Infrared spectra are reported for the polycrystalline compound, isotopically normal and partially deuterated, at temperatures ranging between ca. 80 K and 293 K together with the room temperature Raman spectrum. The assignment of the observed bands was accomplished assuming the existence of one type of pentacyanonitrosylchromate ion in the asymmetric unit, as suggested by the single band found in the NO stretching region of the deuterated anion and in the anhydrous compound. TGA-DTA data are also reported and discussed. The assignments are supported by DFT calculations of the normal modes of vibration of the [Cr(CN)5NO](3-) structure, optimized at the same level of theory.

[The intercontinental schizophrenia outpatient health outcomes study (IC-SOHO): initial 6 month findings of the sample in Latin America]. / Estudio observacional intercontinental de los resultados de salud en pacientes ambulatorios con esquizofrenia (IC-SOHO): hallazgos iniciales de 6 meses de la muestra en Latinoamérica.

The IC-SOHO study was designed to supply information on antipsychotic treatments in the real clinical practice by assessment of a large and diverse sample population with schizophrenia. This document describes the findings of the first 6 months of IC-SOHO in Latin America. To date, this is the largest observational study of its type in this region. In this observational and prospective study, those out-patients with schizophrenia, who require a change or initiation of antipsychotic medication are hospitalized. Effectiveness was evaluated using the Clinical Global Impression-Seriousness (CGI-S) grading scale. Tolerability was assessed by questionnaires on adverse events and weight measurements. Herein, the comparisons between olanzapine (monotherapy), risperidone (monotherapy) and conventional antipsychotics (monotherapy and combined therapy) are presented. As a whole, 7,658 patients participated in the ICSOHO; n=2,671 from 11 countries of Latin America that were included in this report. At 6 months, the proportion of patients who responded to olanzapine was significantly greater than those who responded to risperidone or conventional antipsychotics (p<0.001). Patients from the olanzapine group had greater improvements in all the symptom domains, including general, positive, negative, depressive and cognitive symptoms in comparison with risperidone (p<0.05) or conventional antipsychotics (p < 0.001). Extrapyramidal symptoms (EPS) and tardive dyskinesia (TD) decreased from baseline in the groups treated with olanzapine and risperidone, but increased in the conventional group. The adverse events related with the sexual function were more prominent in the conventional group. Weight gain was observed in each treatment group, although the patients from the olanzapine group had greater weight grain followed by those of risperidone and then by those of conventional antipsychotics. Our findings in this population of the Latin American sample emulate the results of other studies in different samples, where it was found that olanzapine was more effective and better tolerated than risperidone or conventional antipsychotics.

Estudio observacional intercontinental de los resultados de salud en pacientes ambulatorios con esquizofrenia (IC-SOHO): hallazgos iniciales de 6 meses de la muestra en Latinoamérica / The intercontinental schizophrenia outpatient health outcomes study (IC-SOHO): initial 6 month findings of the sample in Latin America

El estudio IC-SOHO se diseñó para aportar información sobre los tratamientos antipsicóticos en la práctica clínica real mediante la evaluación de una población de muestra grande y diversa con esquizofrenia. Este documento describe los hallazgos de los primeros 6 meses del IC-SOHO en Latinoamérica. A la fecha éste es el estudio observacional más grande de su tipo en esta región. En este estudio observacional y prospectivo se ingresaron aquellos pacientes ambulatorios con esquizofrenia que requirieron un cambio o un inicio de medicación antipsicótica. La efectividad se evaluó utilizando la escala de Calificación de Impresión Clínica Global-Gravedad (CGI-S). La tolerabilidad se evaluó mediante cuestionarios de efectos adversos y mediciones de peso. Se presentan aquí las comparaciones entre olanzapina (monoterapia), risperidona (monoterapia) y antipsicóticos convencionales (monoterapia y terapia combinada). En conjunto, participaron 7.658 pacientes en el IC-SOHO; n=2.671 provenientes de 11 países de Latinoamérica se incluyeron en este informe. A los 6 meses la proporción de pacientes que respondieron a la olanzapina fue significativamente mayor que los que respondieron a la risperidona o los antipsicóticos convencionales (p < 0,001). Los pacientes del grupo de olanzapina tuvieron mejorías mayores en todos los dominios de síntomas, incluyendo los síntomas generales, positivos, negativos, depresivos y cognoscitivos, en comparación con la risperidona (p<0,05) o los antipsicóticos convencionales (p<0,001). Los síntomas extrapiramidales (SEP) y la discinesia tardía (DT) disminuyeron desde la línea basal en los grupos tratados con olanzapina y risperidona, pero aumentaron en el grupo convencional. Los efectos adversos relacionados con la función sexual fueron más prominentes en el grupo convencional. Se observó ganancia de peso en cada grupo de tratamiento, aunque los pacientes del grupo de olanzapina aumentaron más de peso, seguidos por los de risperidona y después por los de antipsicóticos convencionales. Nuestros hallazgos en esta población de muestra latinoamericana emulan los resultados de otros estudios en muestras diferentes, donde se encontró que la olanzapina fue más efectiva y mejor tolerada que la risperidona o los antipsicóticos convencionales

The IC-SOHO study was designed to supply information on antipsychotic treatments in the real clinical practice by assessment of a large and diverse sample population with schizophrenia. This document describes the findings of the first 6 months of IC-SOHO in Latin America. To date, this is the largest observational study of its type in this region. In this observational and prospective study, those out-patients with schizophrenia, who require a change or initiation of antipsychotic medication are hospitalized. Effectiveness was evaluated using the Clinical Global Impression- Seriousness (CGI-S) grading scale. Tolerability was assessed by questionnaires on adverse events and weight measurements. Herein, the comparisons between olanzapine (monotherapy), risperidone (monotherapy) and conventional antipsychotics (monotherapy and combined therapy) are presented. As a whole, 7,658 patients participated in the ICSOHO; n=2,671 from 11 countries of Latin America that were included in this report. At 6 months, the proportion of patients who responded to olanzapine was significantly greater than those who responded to risperidone or conventional antipsychotics (p<0.001). Patients from the olanzapine group had greater improvements in all the symptom domains, including general, positive, negative, depressive and cognitive symptoms in comparison with risperidone (p<0.05) or conventional antipsychotics (p < 0.001). Extrapyramidal symptoms (EPS) and tardive dyskinesia (TD) decreased from baseline in the groups treated with olanzapine and risperidone, but increased in the conventional group. The adverse events related with the sexual function were more prominent in the conventional group. Weight gain was observed in each treatment group, although the patients from the olanzapine group had greater weight grain followed by those of risperidone and then by those of conventional antipsychotics. Our findings in this population of the Latin American sample emulate the results of other studies in different samples, where it was found that olanzapine was more effective and better tolerated than risperidone or conventional antipsychotics

Agreement between paramedic-estimated weights and subsequent hospital measurements in adults with out-of-hospital cardiac arrest.

OBJECTIVE: To assess the accuracy of paramedic estimates of adult body weights in cardiac arrest cases. HYPOTHESIS: Paramedics could accurately estimate the weights of out-of-hospital cardiac arrest patients. DESIGN: Retrospective data analysis of a 15-month, multicenter study involving nontraumatic out-of-hospital cardiac arrest patients. Paramedic estimates of body weights were compared to weights measured in the hospital. Patients were included in the analysis only if both a paramedic weight and a measured in-hospital weight were recorded. SETTING: Six urban emergency medical services systems. PARTICIPANTS: The study population included adults with return of spontaneous circulation who subsequently were admitted to the hospital. MEASUREMENTS: Pearson correlation analysis of paramedic-estimated weights and measured weights. RESULTS: Among the 133 study patients, the correlation coefficient (R) for paramedic estimates and the actual measured weight was 0.93. Paramedic estimates of weight were within 10% of the measured weights in 74% of the patients, and within 20% of measured weights in 93% of the patients. CONCLUSION: Paramedic weight estimates correlated well with measured weights.