De-escalated therapy for HR+/HER2+ breast cancer patients with Ki67 response after 2-week letrozole: results of the PerELISA neoadjuvant study.

BACKGROUND: In human epidermal growth factor receptor 2 (HER2+) breast cancers, neoadjuvant trials of chemotherapy plus anti-HER2 treatment consistently showed lower pathologic complete response (pCR) rates in hormone receptor (HR) positive versus negative tumors. The PerELISA study was aimed to evaluate the efficacy of a de-escalated, chemotherapy-free neoadjuvant regimen in HR+/HER2+ breast cancer patients selected on the basis of Ki67 inhibition after 2-week letrozole. PATIENTS AND METHODS: PerELISA is a phase II, multicentric study for postmenopausal patients with HR+/HER2+ operable breast cancer. Patients received 2-week letrozole, and then underwent re-biopsy for Ki67 evaluation. Patients classified as molecular responders (Ki67 relative reduction >20% from baseline) continued letrozole and started trastuzumab-pertuzumab for five cycles. Patients classified as molecular non-responders started weekly paclitaxel for 13 weeks combined with trastuzumab-pertuzumab. Primary aim was breast and axillary pCR. According to a two-stage Simon's design, to reject the null hypothesis, at least 8/43 pCR had to be documented. RESULTS: Sixty-four patients were enrolled, 44 were classified as molecular responders. All these patients completed the assigned treatment with letrozole-trastuzumab-pertuzumab and underwent surgery. A pCR was observed in 9/44 cases (20.5%, 95% confidence interval 11.1% to 34.5%). Among molecular non-responders, 16/17 completed treatment and underwent surgery, with pCR observed in 81.3% of the cases. PAM50 intrinsic subtype was significantly associated with Ki67 response and pCR. Among molecular responders, the pCR rate was significantly higher in HER2-enriched than in other subtypes (45.5% versus 13.8%, P = 0.042). CONCLUSIONS: The primary end point of the study was met, by reaching the pre-specified pCRs. In patients selected using Ki67 reduction after short-term letrozole exposure, a meaningful pCR rate can be achieved without chemotherapy. PAM50 intrinsic subtyping further refines our ability to identify a subset of patients for whom chemotherapy might be spared. EUDRACT NUMBER: 2013-002662-40. CLINICALTRIALS.GOV IDENTIFIER: NCT02411344.

A predictive model of pathologic response based on tumor cellularity and tumor-infiltrating lymphocytes (CelTIL) in HER2-positive breast cancer treated with chemo-free dual HER2 blockade.

Background: The presence of stromal tumor-infiltrating lymphocytes (TILs) is associated with increased pathologic complete response (pCR) and improved outcomes in HER2-positive early-breast cancer (BC) treated with anti-HER2-based chemotherapy. In the absence of chemotherapy, the association of TILs with pCR following anti-HER2 therapy-only is largely unknown. Patients and methods: The PAMELA neoadjuvant trial treated 151 women with HER2-positive BC with lapatinib and trastuzumab [and hormonal therapy if hormone receptor (HR)-positive] for 18 weeks. Percentage of TILs and tumor cellularity were determined at baseline (N = 148) and at day 15 (D15) of treatment (N = 134). Associations of TILs and tumor cellularity with pCR in the breast were evaluated. A combined score based on tumor cellularity and TILs (CelTIL) measured at D15 was derived in PAMELA, and validated in D15 samples from 65 patients with HER2-positive disease recruited in the LPT109096 neoadjuvant trial, where anti-HER2 therapy-only was administer for 2 weeks, then standard chemotherapy was added for 24 weeks. Results: In PAMELA, baseline and D15 TILs were significantly associated with pCR in univariate analysis. In multivariable analysis, D15 TILs, but not baseline TILs, were significantly associated with pCR. At D15, TILs and tumor cellularity were found independently associated with pCR. A combined score (CelTIL) taking into account both variables was derived. CelTIL at D15 as a continuous variable was significantly associated with pCR, and patients with CelTIL-low and CelTIL-high scores had a pCR rate of 0% and 33%, respectively. In LPT109096, CelTIL at D15 was found associated with pCR both as a continuous variable and as group categories using a pre-defined cut-off (75.0% versus 33.3%). Conclusions: On-treatment TILs, but not baseline TILs, are independently associated with response following anti-HER2 therapy-only. A combined score of TILs and tumor cellularity measured at D15 provides independent predictive information upon completion of neoadjuvant anti-HER2-based therapy. Clinical trial number: NCT01973660.

Association between PD1 mRNA and response to anti-PD1 monotherapy across multiple cancer types.

Background: We hypothesized that the abundance of PD1 mRNA in tumor samples might explain the differences in overall response rates (ORR) observed following anti-PD1 monotherapy across cancer types. Patients and methods: RNASeqv2 data from 10 078 tumor samples representing 34 different cancer types was analyzed from TCGA. Eighteen immune-related gene signatures and 547 immune-related genes, including PD1, were explored. Correlations between each gene/signature and ORRs reported in the literature following anti-PD1 monotherapy were calculated. To translate the in silico findings to the clinical setting, we analyzed the expression of PD1 mRNA using the nCounter platform in 773 formalin-fixed paraffin embedded (FFPE) tumor samples across 17 cancer types. To test the direct relationship between PD1 mRNA, PDL1 immunohistochemistry (IHC), stromal tumor-infiltrating lymphocytes (sTILs) and ORR, we evaluated an independent FFPE-based dataset of 117 patients with advanced disease treated with anti-PD1 monotherapy. Results: In pan-cancer TCGA, PD1 mRNA expression was found strongly correlated (r > 0.80) with CD8 T-cell genes and signatures and the proportion of PD1 mRNA-high tumors (80th percentile) within a given cancer type was variable (0%-84%). Strikingly, the PD1-high proportions across cancer types were found strongly correlated (r = 0.91) with the ORR following anti-PD1 monotherapy reported in the literature. Lower correlations were found with other immune-related genes/signatures, including PDL1. Using the same population-based cutoff (80th percentile), similar proportions of PD1-high disease in a given cancer type were identified in our in-house 773 tumor dataset as compared with TCGA. Finally, the pre-established PD1 mRNA FFPE-based cutoff was found significantly associated with anti-PD1 response in 117 patients with advanced disease (PD1-high 51.5%, PD1-intermediate 26.6% and PD1-low 15.0%; odds ratio between PD1-high and PD1-intermediate/low = 8.31; P < 0.001). In this same dataset, PDL1 tumor expression by IHC or percentage of sTILs was not found associated with response. Conclusions: Our study provides a clinically applicable assay that links PD1 mRNA abundance, activated CD8 T-cells and anti-PD1 efficacy.

Unraveling the clinicopathological and molecular changes induced by neoadjuvant chemotherapy and endocrine therapy in hormone receptor-positive/HER2-low and HER2-0 breast cancer.

BACKGROUND: The characterization and comparison of gene expression and intrinsic subtype (IS) changes induced by neoadjuvant chemotherapy (NACT) and endocrine therapy in hormone receptor-positive (HR+)/human epidermal growth factor receptor 2 (HER2)-low versus HR+/HER2-0 breast cancer (BC) has not been conducted so far. Most evidence on the association of HER2 status with pathologic responses and prognosis in HR+/HER2-negative BC is controversial and restricted to NACT-treated disease. Similarly, a temporal heterogeneity in HER2 status has been described only with NACT. METHODS: We retrospectively recruited a consecutive cohort of 186 patients with stage I-IIIB HR+/HER2-negative BC treated with neoadjuvant therapy (NAT). Available diagnostic biopsies and surgical samples were characterized for main pathological features, PAM50 IS and ROR-P score, and gene expression. Associations with pathologic complete response, residual cancer burden-0/I, event-free survival (EFS) and overall survival (OS) based on HER2 status were assessed. Pre/post pathologic/molecular changes were analyzed in matched samples. RESULTS: The HER2-low (62.9%) and HER2-0 (37.1%) cohorts did not differ significantly in main baseline features, treatments administered, breast-conserving surgery, pathologic complete response and residual cancer burden-0/I rates, EFS, and OS. NAT induced, regardless of HER2 status, a significant reduction of estrogen receptor/progesterone receptor and Ki67 levels, a down-regulation of PAM50 proliferation- and luminal-related genes/signatures, an up-regulation of selected immune genes, and a shift towards less aggressive IS and lower ROR-P. Moreover, 25% of HER2-0 changed to HER2-low and 34% HER2-low became HER2-0. HER2 shifts were significant after NACT (P < 0.001), not neoadjuvant endocrine therapy (P = 0.063), with consistent ERBB2 mRNA level dynamics. HER2 changes were not associated with EFS/OS. CONCLUSIONS: HER2-low and HER2-0 status change after NAT in ∼30% of cases, mostly after NACT. Targeted adjuvant strategies should be investigated accordingly. Molecular downstaging with current chemo/endocrine agents and immunotherapy should not rely on HER2 immunohistochemical levels in HR+/HER2-negative BC. Instead, HER2-low-targeted approaches should be explored to pursue more effective and/or less toxic dimensional downstaging.

Analytical validation of HER2DX genomic test for early-stage HER2-positive breast cancer.

BACKGROUND: HER2DX, a multianalyte genomic test, has been clinically validated to predict breast cancer recurrence risk (relapse risk score), the probability of achieving pathological complete response post-neoadjuvant therapy (pCR likelihood score), and individual ERBB2 messenger RNA (mRNA) expression levels in patients with early-stage human epidermal growth factor receptor 2 (HER2)-positive breast cancer. This study delves into the comprehensive analysis of HER2DX's analytical performance. MATERIALS AND METHODS: Precision and reproducibility of HER2DX risk, pCR, and ERBB2 mRNA scores were assessed within and between laboratories using formalin-fixed paraffin-embedded (FFPE) tumor tissues and purified RNA. Robustness was appraised by analyzing the impact of tumor cell content and protocol variations including different instruments, reagent lots, and different RNA extraction kits. Variability was evaluated across intratumor biopsies and genomic platforms [RNA sequencing (RNAseq) versus nCounter], and according to protocol variations. RESULTS: Precision analysis of 10 FFPE tumor samples yielded a maximal standard error of 0.94 across HER2DX scores (1-99 scale). High reproducibility of HER2DX scores across 29 FFPE tumors and 20 RNAs between laboratories was evident (correlation coefficients >0.98). The probability of identifying score differences >5 units was ≤5.2%. No significant variability emerged based on platform instruments, reagent lots, RNA extraction kits, or TagSet thaw/freeze cycles. Moreover, HER2DX displayed robustness at low tumor cell content (10%). Intratumor variability across 212 biopsies (106 tumors) was <4.0%. Concordance between HER2DX scores from 30 RNAs on RNAseq and nCounter platforms exceeded 90.0% (Cohen's κ coefficients >0.80). CONCLUSIONS: The HER2DX assay is highly reproducible and robust for the quantification of recurrence risk, pCR likelihood, and ERBB2 mRNA expression in early-stage HER2-positive breast cancer.

Identification of cell surface targets for CAR-T cell therapies and antibody-drug conjugates in breast cancer.

BACKGROUND: Two promising therapeutic strategies in oncology are chimeric antigen receptor-T cell (CAR-T) therapies and antibody-drug conjugates (ADCs). To be effective and safe, these immunotherapies require surface antigens to be sufficiently expressed in tumors and less or not expressed in normal tissues. To identify new targets for ADCs and CAR-T specifically targeting breast cancer (BC) molecular and pathology-based subtypes, we propose a novel in silico strategy based on multiple publicly available datasets and provide a comprehensive explanation of the workflow for a further implementation. METHODS: We carried out differential gene expression analyses on The Cancer Genome Atlas BC RNA-sequencing data to identify BC subtype-specific upregulated genes. To fully explain the proposed target-discovering methodology, as proof of concept, we selected the 200 most upregulated genes for each subtype and undertook a comprehensive analysis of their protein expression in BC and normal tissues through several publicly available databases to identify the potentially safest and viable targets. RESULTS: We identified 36 potentially suitable and subtype-specific tumor surface antigens (TSAs), including fibroblast growth factor receptor-4 (FGFR4), carcinoembryonic antigen-related cell adhesion molecule 6 (CEACAM6), GDNF family receptor alpha 1 (GFRA1), integrin beta-6 (ITGB6) and ectonucleotide pyrophosphatase/phosphodiesterase 1 (ENPP1). We also identified 63 potential TSA pairs that might be appropriate for co-targeting strategies. Finally, we validated subtype specificity in a cohort of our patients, multiple BC cell lines and the METABRIC database. CONCLUSIONS: Overall, our in silico analysis provides a framework to identify novel and specific TSAs for the development of new CAR-T and antibody-based therapies in BC.

Immune microenvironment characterisation and dynamics during anti-HER2-based neoadjuvant treatment in HER2-positive breast cancer.

Despite their recognised role in HER2-positive (HER2+) breast cancer (BC), the composition, localisation and functional orientation of immune cells within tumour microenvironment, as well as its dynamics during anti-HER2 treatment, is largely unknown. We here investigate changes in tumour-immune contexture, as assessed by stromal tumour-infiltrating lymphocytes (sTILs) and by multiplexed spatial cellular phenotyping, during treatment with lapatinib-trastuzumab in HER2+ BC patients (PAMELA trial). Moreover, we evaluate the relationship of tumour-immune contexture with hormone receptor status, intrinsic subtype and immune-related gene expression. sTIL levels increase after 2 weeks of HER2 blockade in HR-negative disease and HER2-enriched subtype. This is linked to a concomitant increase in cell density of all four immune subpopulations (CD3+, CD4+, CD8+, Foxp3+). Moreover, immune contexture analysis showed that immune cells spatially interacting with tumour cells have the strongest association with response to anti-HER2 treatment. Subsequently, sTILs consistently decrease at the surgery in patients achieving pathologic complete response, whereas most residual tumours at surgery remain inflamed, possibly reflecting a progressive loss of function of T cells. Understanding the features of the resulting tumour immunosuppressive microenvironment has crucial implications for the design of new strategies to de-escalate or escalate systemic therapy in early-stage HER2+ BC.

Level statistics of words: finding keywords in literary texts and symbolic sequences.

Using a generalization of the level statistics analysis of quantum disordered systems, we present an approach able to extract automatically keywords in literary texts. Our approach takes into account not only the frequencies of the words present in the text but also their spatial distribution along the text, and is based on the fact that relevant words are significantly clustered (i.e., they self-attract each other), while irrelevant words are distributed randomly in the text. Since a reference corpus is not needed, our approach is especially suitable for single documents for which no a priori information is available. In addition, we show that our method works also in generic symbolic sequences (continuous texts without spaces), thus suggesting its general applicability.

Forward signaling mediated by ephrin-B3 prevents contralateral corticospinal axons from recrossing the spinal cord midline.

To investigate Eph-ephrin bidirectional signaling, a series of mutations were generated in the ephrin-B3 locus. The absence of both forward and reverse signaling resulted in mice with mirror movements as typified by a hopping locomotion. The corticospinal tract was defective as axons failed to respect the midline boundary of the spinal cord and bilaterally innervated both contralateral and ipsilateral motor neuron populations. A second mutation that expresses a truncated ephrin-B3 protein lacking its cytoplasmic domain did not lead to hopping, indicating that reverse signaling is not required for corticospinal innervation. Ephrin-B3 is concentrated at the spinal cord midline, while one of its receptors, EphA4, is expressed in postnatal corticospinal neurons as their fibers pathfind down the contralateral spinal cord. Our data indicate ephrin-B3 functions as a midline-anchored repellent to stimulate forward signaling in EphA4-expressing axons.

Effect of magnetic ion exchange (MIEX®) on removal of emerging organic contaminants.

In this study, the removal of nine emerging organic contaminants was investigated by using anion exchange resins. The selected compounds were carbamazepine, atrazine, simazine, estrone, bisphenol A, methylparaben, ethylparaben, propylparaben and butylparaben. Two different magnetic anionic exchanger resins were tested: MIEX® DOC and MIEX® GOLD. The optimal resin dose (40 mL/L) and contact time (20 min) had been previously determined. Once these optimum parameters were set, the effect of the initial concentration of contaminants on the removal efficiency of the contaminants by the resins was studied. The study was carried out using mono and multicomponent systems, with distilled water and natural waters, to which contaminants had been previously added, in order to evaluate the competitive and matrix effects. Results showed that the average removal percentages obtained with the MIEX® DOC resin were: 51%, 61%, 68% and 80% for methyl-, ethyl-, propyl-, and butylparaben, respectively. For bisphenol A the result was similar, i.e., 66%, whereas for the rest of the compounds studied, removal efficiencies lower than 15% were obtained. The MIEX® GOLD resin achieved lower elimination rates than the MIEX® DOC resin in all cases.

Identifying characteristic scales in the human genome.

The scale-free, long-range correlations detected in DNA sequences contrast with characteristic lengths of genomic elements, being particularly incompatible with the isochores (long, homogeneous DNA segments). By computing the local behavior of the scaling exponent alpha of detrended fluctuation analysis (DFA), we discriminate between sequences with and without true scaling, and we find that no single scaling exists in the human genome. Instead, human chromosomes show a common compositional structure with two characteristic scales, the large one corresponding to the isochores and the other to small and medium scale genomic elements.

Polycyclic aromatic hydrocarbons (PAHs) in human milk from Italian women: influence of cigarette smoking and residential area.

The presence of PAH in breast milk collected from 32 smoking and non-smoking lactating women, residing in urban and rural areas of Tuscany (Italy) was investigated. The results indicated a significant contribution of tobacco smoke to the PAH contamination of milk: the condensate contained in the cigarettes smoked daily by each subject was strongly related with the polynuclear hydrocarbon content (R(2)=0.92, P<0.005). An experiment carried out under controlled exposure conditions to cigarette smoke allowed to demonstrate that individual metabolic activity and smoking habits affect the PAH concentration in milk samples. Mothers living in rural environments showed significantly lower PAH concentrations than those observed in urban subjects. The risk evaluation due to PAH ingestion via breast milk was assessed on the basis of the acceptable daily intake of Benzo(a)pyrene in drinking water, evidencing that a hazard cannot be excluded for heavy smokers residing in urban areas.

Determinación de las características funcionales de los nódulos tiroideos mediante imágenes de medicina nuclear en pacientes que acudieron al Instituto de Investigaciones en Ciencias de la Salud - UNA, en el periodo 2016-2019 / Determination of the functional characteristics of thyroid nodules through nuclear medicine images in patients, who attended the Instituto de Investigaciones en Ciencias de la Salud - UNA, in the period 2016-2019

nuclear permiten visualizar la estructura y función de un órgano, tejido, hueso o sistema dentro del cuerpo, entre ellos la glándula tiroidea, la cual puede presentar un nódulo tiroideo y este es importante diagnosticar por su potencial malignidad. Objetivo: Determinar las características funcionales de los nódulos tiroideos en 183 pacientes que asistieron al Servicio de Medicina Nuclear del Instituto de Investigaciones en Ciencias de la Salud - UNA durante el período de noviembre del 2016 - noviembre del 2019. Metodología: Fue utilizada la gammacámara SPECT doble cabezal, siguiendo protocolo estandarizado, con administración de pertecnetato de sodio. Resultados: Se observó que la gran mayoría de los pacientes corresponden al sexo femenino 89%, así como que el 64% provienen de la capital y del departamento central; las patologías detectadas corresponden a bocio en 82%, 15% a nódulos (hipercaptantes, hipocaptantes y autónomos) y casos particulares de tiroiditis, tiroides ectópica y adenoma tóxico. La medicina nuclear está directamente involucrada tanto en el diagnóstico como en el tratamiento de la enfermedad tiroidea, por lo que se requiere una comprensión de la fisiopatología y el manejo de los trastornos de la tiroides, de manera a que las políticas de salud pública sean implementadas para el fortalecimiento de la lucha contra estas enfermedades. Conclusión: El trabajo de investigación realizado comprobó que un mayoritario porcentaje de participantes de sexo femenino (89%), con enfermedades tiroideas asistieron al IICS/UNA, y que la frecuencia de bocio entre la totalidad de pacientes que participaron de este estudio fué de 151/183 (82%), los pacientes provenían en su mayoría de hospitales de referencia del departamento Central y coinciden con la localización geográfica de los domicilios de los pacientes, quienes provenían principalmente del mismo departamento Central 118/183 (64%), no obstante también participaron del trabajo de investigación pacientes provenientes de otros departamentos del país. Palabras claves: Diagnóstico, medicina nuclear, tiroides, tecnología nuclear en salud, SPECT

Introduction: The images obtained by nuclear medicine allow to visualize the structure and function of an organ, tissue, bone or system inside the body, including the thyroid gland, which can present a thyroid nodule and this is important to diagnose for its potential malignancy. Objective: To determine the functional characteristics of the thyroid nodules were determined in 183 patients, who attended the Nuclear Medicine Service at the Instituto de Investigaciones en Ciencias de la Salud during the period of November 2016 - November 2019. Methodology: It was used the dual head SPECT gamma camera following a standardized protocol with administration of sodium pertechnetate. Result: It was observed that most of the patients corresponded to the female sex 89 %, as well as that 64% came from the capital and the central department. The pathologies detected corresponded to goiter in 82%, 15% to nodules (hypercaptant, hypocaptant and autonoms) and particular cases of thyroiditis, ectopic thyroid and toxic adenoma. Nuclear medicine is directly involved both in the diagnosis and in the treatment of thyroid disease, therefore it requires an understanding of the pathophysiology and management of thyroid disorders in order that public health policies are implemented for strengthening the fight against these diseases. Conclusion: The research carried out showed that a majority percentage of female participants (89%), with thyroid diseases attended IICS / UNA, and the frequency of goiter among all patients who participated in this study was 151/183 ( 82%), the patients mostly they came the reference hospitals in the Central department and coincide with the geographic location of the patients' homes, who tested mainly from the same Central department 118/183 (64%), also participate in the research work patients from other departments of the country. Keywords: Diagnosis, nuclear medicine, thyroid, health technology, SPECT.

Isochore chromosome maps of eukaryotic genomes.

Analytical DNA ultracentrifugation revealed that eukaryotic genomes are mosaics of isochores: long DNA segments (>>300 kb on average) relatively homogeneous in G+C. Important genome features are dependent on this isochore structure, e.g. genes are found predominantly in the GC-richest isochore classes. However, no reliable method is available to rigorously partition the genome sequence into relatively homogeneous regions of different composition, thereby revealing the isochore structure of chromosomes at the sequence level. Homogeneous regions are currently ascertained by plain statistics on moving windows of arbitrary length, or simply by eye on G+C plots. On the contrary, the entropic segmentation method is able to divide a DNA sequence into relatively homogeneous, statistically significant domains. An early version of this algorithm only produced domains having an average length far below the typical isochore size. Here we show that an improved segmentation method, specifically intended to determine the most statistically significant partition of the sequence at each scale, is able to identify the boundaries between long homogeneous genome regions displaying the typical features of isochores. The algorithm precisely locates classes II and III of the human major histocompatibility complex region, two well-characterized isochores at the sequence level, the boundary between them being the first isochore boundary experimentally characterized at the sequence level. The analysis is then extended to a collection of human large contigs. The relatively homogeneous regions we find show many of the features (G+C range, relative proportion of isochore classes, size distribution, and relationship with gene density) of the isochores identified through DNA centrifugation. Isochore chromosome maps, with many potential applications in genomics, are then drawn for all the completely sequenced eukaryotic genomes available.

A simple and species-independent coding measure.

We present a coding measure which is based on the statistical properties of the stop codons, and that is able to estimate accurately the variation of coding content along an anonymous sequence. As the stop codons play the same role in all the genomes (with very few exceptions) the measure turns out to be species-independent. We show results both for prokaryotic and for eukaryotic genomes, indicating, first, the accuracy of the measure, and, second, that better prediction is achieved if the measure is applied on homogeneous, isochore-like sequences than if it is applied following the standard moving window approach. Finally, we discuss on some of the possible applications of the measure.

Study of statistical correlations in DNA sequences.

Here we present a study of statistical correlations among different positions in DNA sequences and their implications by directly using the autocorrelation function. Such an analysis is possible now because of the availability of large sequences or even complete genomes of many organisms. After describing the way in which the autocorrelation function can be applied to DNA-sequence analysis, we show that long-range correlations, implying scale independence, appear in several bacterial genomes as well as in long human chromosome contigs. The source for such correlations in bacteria, which may extend up to 60 kb in Bacillus subtilis, may be related to massive lateral transfer of compositionally biased genes from other genomes. In the human genome, correlations extend for more than five decades and may be related to the evolution of the 'neogenome', a modern evolutionary acquisition composed by GC-rich isochores displaying long-range correlations and scale invariance.

Decrease of cyclin D1 in the human lung adenocarcinoma cell line A-427 by 7-hydroxycoumarin.

Coumarin in vivo has antitumor activity in various types of cancer. In vitro, coumarin and 7-hydroxycoumarin, its major biotransformation product in humans, inhibit the proliferation of several human tumor cell lines. The molecular mechanisms of these effects are unknown. To gain information about these mechanisms, we studied the effects of coumarin and 7-hydroxycoumarin in the human lung adenocarcinoma cell line A-427 on the inhibition of: (i) cell proliferation; (ii) cell cycle progression; and (iii) expression of cyclins D1, E and A. The inhibitory concentrations 50 (IC(50)) of both compounds were estimated by cytostatic assays of tetrazolium (MTT) reduction. The effects on cell cycle progression were assayed with propidium iodide and BrdU using DNA histograms and multiparametric flow cytometry. The percentages of cells expressing cyclins D1, E, and A were estimated by means of bivariate flow cytometry using propidium iodide, and FITC-conjugated monoclonal antibodies for each cyclin. The IC(50) (+/-S.E.M. n=3) of 7-hydroxycoumarin and coumarin at 72 h exposure, were 100+/-4.8 and 257+/-8.8 microg/ml, respectively. 7-Hydroxycoumarin at the concentration of 160 microg/ml (1 mM), inhibited the G(1)/S transition of the cell cycle, an action consistent with the cytostatic effect. No significant decreases of cyclins E and A were observed. In contrast, cyclin D1 significantly decreased, which appears to indicate an action of 7-hydroxycoumarin in early events of phase G(1). However, messenger RNA of cyclin D1, assayed by RT-PCR, did not change. This suggests a posttranscriptional effect. The effects of coumarin were not significant. Cyclin D1 is overexpressed in many types of cancer, and its inhibition has been proposed as a pharmacological and therapeutic target for novel antitumor agents. Knowledge of the decrease of cyclin D1 by 7-hydroxycoumarin may lead to its use in cancer therapy, as well as to the development of more active compounds.

Bilirubin photoisomerization products in serum and urine from a Crigler-Najjar type I patient treated by phototherapy.

The relative compositions of the photoisomers of bilirubin-1X alpha (4Z, 15Z-bilirubin) in serum and urine of a patient with Crigler-Najjar type I syndrome treated by phototherapy are reported. High-performance liquid chromatography analysis reveals the presence of high serum levels of the configurational bilirubin photoisomer (4Z,15E-bilirubin) before the beginning of phototherapy (between 12 and 16% of the total bilirubin). The configurational photoisomer value increases during phototherapy with blue fluorescent lamps up to a photoequilibrium of about 25%, similar to that obtained in a bilirubin solution in vitro irradiated by the same lamps. This evidence suggests an inefficient serum excretion of the 4Z,15E-bilirubin. Indeed, its average half-life in serum of the Crigler-Najjar patient is found to be about 8 h. No detectable traces of the bilirubin structural isomer, lumirubin, are found in the serum. On the other hand, lumirubin represents the dominant bilirubin isomer excreted in the urine, as both 15Z and 15E configurations. Smaller amounts of 4Z,15E-bilirubin, 4E,15Z-bilirubin and native 4Z,15Z-bilirubin are observed in urine. The presence in urine of 4Z,15Z-bilirubin is probably due to a fast reversion of the configurational photoisomers to their native form. The half-life of the configurational photoisomers in urine kept at 38 degrees C is found to be of the order of a few minutes. Our study indicates that in Crigler-Najjar type I patients, mechanisms exist to excrete all bilirubin photoisomers. The lumirubin pathway seems to contribute markedly to bilirubin excretion in the urine, as occurs in jaundiced babies under phototherapy. However, the contribution of configurational isomers cannot be neglected.

[Residual symptoms in elderly patients with depression]. / Síntomas residuales en el anciano con depresión.

OBJECTIVES: To look for residual symptoms in patients older than 65 years diagnosed with depression, and to find out if there are more residual symptoms in relation to physical comorbidity and the number of visits made to a Primary Care Centre. METHODS: A descriptive, randomised and cross-sectional multicentre study, with a sample of patients attending 3 urban Health Primary Care Centres. Patients older than 65 years with a diagnosis of depression and who had received more than 3 months treatment were selected. Data was collected by means of questionnaires during personal interviews, which included universal data, time of onset, treatment recurrence, comorbidity, and residual symptoms. The Yesavage questionnaire, EQ-5D and VAS were also used. RESULTS: Of 100 patients studied, 99% had residual symptoms, with 22% with 4 symptoms (from 10 asked). The most frequent symptom was the sadness (68%), followed by pain (55%), and loss of energy (55%). CONCLUSIONS: The prevalence of depression found in our study was 13.8%, and a 99% had at least one residual symptom. Patients who lived as a couple and men (who performed more recreational activities) had a lower number of residual symptoms. No relationship was found between comorbidity and frequent use of Primary Care Centres with a higher number of residuals symptoms. The majority of patients suffered chronic depression (> 2 years). Benzodiazepines only were used as treatment in 18% of the patients.

Cobertura Universal de Servicios Diagnósticos a través de la Telemedicina / Universal Coverage of Diagnostic Services through Telemedicine

Con la Telemedicina pueden desarrollarse sistemas de telediagnóstico ventajosos para mejorar la atención de la salud de poblaciones remotas que no tienen acceso a especialistas. Este estudio realizado por la Unidad de Telemedicina del Ministerio de Salud Pública y Bienestar Social (MSPBS) en colaboración con el Dpto. de Ingeniería Biomédica e Imágenes del Instituto de Investigaciones en Ciencias (IICS-UNA) y la Universidad del País Vasco (UPV/EHU) sirvió para evaluar la utilidad de un sistema de telediagnóstico para la cobertura universal en la salud pública. Para el efecto se analizaron los resultados obtenidos por el sistema de telediagnóstico implementado en 56 hospitales del MSPBS. En dicho sentido se analizaron 293.142 diagnósticos remotos realizados entre enero del 2014 y septiembre de 2017. Del total, el 37,29 % (109.311) correspondieron a estudios de tomografía, 61,44 % (180.108) a electrocardiografía (ECG), 1,26 % (3.704) a electroencefalografía (EEG) y 0,01 % (19) a ecografía. No se observaron diferencias significativas entre el diagnóstico remoto y el diagnóstico "cara a cara". Con el diagnostico remoto se logró una reducción del coste que supone un beneficio importante para cada ciudadano del interior del país. Los resultados obtenidos evidencian que la telemedicina puede contribuir para mejorar significativamente la cobertura universal de los servicios diagnósticos y programas de salud, maximizando el tiempo del profesional y su productividad, aumentando el acceso y la equidad, y disminuyendo los costos. Sin embargo antes de realizar su implementación sistemática se deberá realizar una contextualización con el perfil epidemiológico regional. Palabras claves: Telemedicina; Cobertura Universal; Telediagnóstico; TICs en Salud; Innovación Tecnológica.

Through the telemedicine, advantageous telediagnostic systems can be developed to improve the health care of remote populations that don`t have access to specialists. This study was carried out by the Telemedicine Unit of the Ministry of Public Health and Social Welfare (MSPBS) in collaboration with the Department of Biomedical Engineering and Imaging of the Health Science Research Institute (IICS-UNA) and the University of the Basque Country (UPV / EHU) to evaluate the utility of a telediagnostic system for universal coverage in public health. For this purpose, the results obtained by the telediagnosis system implemented in 56 MSPBS hospitals were analyzed. In that sense, 293,142 remote diagnoses were performed between January 2014 and September 2017. Of the total, 37.29% (109,311) corresponded to tomography studies, 61.44% (180.108) to electrocardiography (ECG), 1.26% (3,704) to electroencephalography (EEG) and 0.01% (19) to ultrasound. There were no significant differences between the remote and the "face to face" diagnosis. With the remote diagnosis a reduction of the cost was obtained, that supposes an important benefit for each citizen of the interior of the country. The results show that the telemedicine can contribute to improve significantly the universal coverage of diagnostic services and health programs, maximizing professional time and productivity, increasing access and equity, and reducing costs. However, before carrying out its systematic implementation, a contextualization with the regional epidemiological profile must be performed. Key words: Telemedicine; Universal Coverage; Telediagnosis; ICT in Health; Technological Innovation.