Sensitive, non-immunogenic in vivo imaging of cancer metastases and immunotherapy response.

Non-invasive imaging of tumors expressing reporter transgenes is a popular preclinical method for studying tumor development and response to therapy in vivo due to its ability to distinguish signal from tumors over background noise. However, the utilized transgenes, such as firefly luciferase, are immunogenic and, therefore, impact results when expressed in immune-competent hosts. This represents an important limitation, given that cancer immunology and immunotherapy are currently among the most impactful areas of research and therapeutic development. Here we present a non-immunogenic preclinical tumor imaging approach. Based on the expression of murine sodium iodide symporter (mNIS), it facilitates sensitive, non-invasive detection of syngeneic tumor cells in immune-competent tumor models without additional immunogenicity arising from exogenous transgenic protein or selection marker expression. NIS-expressing tumor cells internalize the gamma-emitting [99mTc]pertechnetate ion and so can be detected by SPECT (single photon emission computed tomography). Using a mouse model of pancreatic ductal adenocarcinoma hepatic metastases in immune-competent C57BL/6 mice, we demonstrate that the technique enables the detection of very early metastatic lesions and longitudinal assessment of immunotherapy responses using precise and quantifiable whole-body SPECT/CT imaging.

A minimally invasive approach to pilonidal disease with endoscopic pilonidal sinus treatment (EPSiT): a single-center case series with long-term results.

BACKGROUND: Minimally invasive techniques for treating pilonidal disease are safe and effective alternatives to conventional surgery, with improved recovery time, cosmetic results, and pain control. The aim of this study was to evaluate the 5-year surgical outcomes of a single-center case series treated with endoscopic pilonidal sinus treatment (EPSiT). METHODS: We conducted a retrospective single-center analysis of all patients treated with EPSiT, by a single surgical team, from March 2015 to December 2019, for primary or recurrent pilonidal disease. The primary outcomes were recurrence, persistence and treatment failure. The secondary outcomes were postoperative pain, painkiller use, time off work, satisfaction, complications, wound healing time, time to persistence or recurrence. RESULTS: Forty-two patients underwent 46 EPSiT procedures [34 males, 8 females, median age 25 (IQR 13.75) years] for primary (47.8%) or recurrent pilonidal disease (52.2%). All patients completed the follow-up [median 62 (IQR 43) months]. The single procedure healing rate was 76.1%. The healing rate for the first procedures plus the second EPSiT procedure (performed in 4 cases) was 83.3%. Among the 46 EPSiT procedures, we recorded six cases of persistence (13.0%) and five cases of recurrence (10.9%) The median operative time was 32.5 (IQR 18.75) minutes, the median pain score (visual analog scale) in week 1 was 2 (IQR 2), and the median time off work was 4 (IQR 2) days. Four patients (8.7%) experienced complications: serosanguineous (n = 2) or seropurulent discharge (n = 2). The satisfaction rate was 95.7%. CONCLUSIONS: In our experience, EPSiT is safe, well accepted. and associated with a low level of postoperative pain, short hospitalization, short time off work, as well as optimal cosmetic results. Its failure rate is similar to that of excisional surgery.

The Effects of Graded Levels of Calorie Restriction: XVI. Metabolomic Changes in the Cerebellum Indicate Activation of Hypothalamocerebellar Connections Driven by Hunger Responses.

Calorie restriction (CR) remains the most robust intervention to extend life span and improve healthspan. Though the cerebellum is more commonly associated with motor control, it has strong links with the hypothalamus and is thought to be associated with nutritional regulation and adiposity. Using a global mass spectrometry-based metabolomics approach, we identified 756 metabolites that were significantly differentially expressed in the cerebellar region of the brain of C57BL/6J mice, fed graded levels of CR (10, 20, 30, and 40 CR) compared to mice fed ad libitum for 12 hours a day. Pathway enrichment indicated changes in the pathways of adenosine and guanine (which are precursors of DNA production), aromatic amino acids (tyrosine, phenylalanine, and tryptophan) and the sulfur-containing amino acid methionine. We also saw increases in the tricarboxylic acid cycle (TCA) cycle, electron donor, and dopamine and histamine pathways. In particular, changes in l-histidine and homocarnosine correlated positively with the level of CR and food anticipatory activity and negatively with insulin and body temperature. Several metabolic and pathway changes acted against changes seen in age-associated neurodegenerative disorders, including increases in the TCA cycle and reduced l-proline. Carnitine metabolites contributed to discrimination between CR groups, which corroborates previous work in the liver and plasma. These results indicate the conservation of certain aspects of metabolism across tissues with CR. Moreover, this is the first study to indicate CR alters the cerebellar metabolome, and does so in a graded fashion, after only a short period of restriction.

PIM Kinase Inhibitors Block the Growth of Primary T-cell Acute Lymphoblastic Leukemia: Resistance Pathways Identified by Network Modeling Analysis.

Despite significant progress in understanding the genetic landscape of T-cell acute lymphoblastic leukemia (T-ALL), the discovery of novel therapeutic targets has been difficult. Our results demonstrate that the levels of PIM1 protein kinase is elevated in early T-cell precursor ALL (ETP-ALL) but not in mature T-ALL primary samples. Small-molecule PIM inhibitor (PIMi) treatment decreases leukemia burden in ETP-ALL. However, treatment of animals carrying ETP-ALL with PIMi was not curative. To model other pathways that could be targeted to complement PIMi activity, HSB-2 cells, previously characterized as a PIMi-sensitive T-ALL cell line, were grown in increasing doses of PIMi. Gene set enrichment analysis of RNA sequencing data and functional enrichment of network modules demonstrated that the HOXA9, mTOR, MYC, NFκB, and PI3K-AKT pathways were activated in HSB-2 cells after long-term PIM inhibition. Reverse phase protein array-based pathway activation mapping demonstrated alterations in the mTOR, PI3K-AKT, and NFκB pathways, as well. PIMi-tolerant HSB-2 cells contained phosphorylated RelA-S536 consistent with activation of the NFκB pathway. The combination of NFκB and PIMis markedly reduced the proliferation in PIMi-resistant leukemic cells showing that this pathway plays an important role in driving the growth of T-ALL. Together these results demonstrate key pathways that are activated when HSB-2 cell line develop resistance to PIMi and suggest pathways that can be rationally targeted in combination with PIM kinases to inhibit T-ALL growth.

Acceso de mujeres en embarazo, parto y post parto a servicios de salud según clase social / Access of women in pregnancy, childbirth and postpartum to health services according to social class

Resumen Objetivo: comprender el acceso que tienen las mujeres en situación de embarazo, parto y post parto a los servicios de salud según la clase social en Bogotá (Colombia). Métodos: estudio cualitativo. Perspectiva hermenéutica crítica. Método etnografía crítica. Muestreo teórico. Análisis por triangulación en Atlas Ti. Participaron 9 mujeres y 8 profesionales de la salud. Se realizaron 38 entrevistas a profundidad durante 13 meses y 62 acompañamientos a las maternas en las actividades de control prenatal, vacunación, trabajo de parto, consulta post parto, exámenes de seguimiento, curso Psicoprofiláctico, hospitalización y sala de espera, tanto en servicios públicos como privados. Resultados: existen desigualdades según la clase social en el acceso que tienen las mujeres a los servicios de salud en los siguientes aspectos: acceso a servicio a especialistas, hacer la fila para esperar la atención, la disponibilidad de citas y agenda para programar la cita, perder la cita habiendo llegado al servicio, la prioridad que le dan las instituciones a las maternas, madrugar para conseguir atención, pedir la cita, las condiciones de la espera y elegir la clínica o el personal. Conclusiones: los anteriores aspectos se intensifican en clases sociales con menos ventajas. Se requiere disminuir las desigualdades sociales para disminuir las inequidades en salud.

Abstract Objective: to understand the access that women in pregnancy, childbirth and postpartum have to health services according to social class in Bogotá (Colombia). Methods: qualitative study. Critical hermeneutical perspective. Critical ethnography method. Theoretical sampling. Analysis by triangulation in Atlas Ti. 9 women and 8 health professionals participated. 38 in-depth interviews were carried out during 13 months and 62 accompaniments to the maternal in the activities of prenatal control, vaccination, labor, postpartum consultation, follow-up examinations, Psychoprophylactic course, hospitalization and waiting room, both in public services as private. Results: there are inequalities according to social class in the access that women have to health services in the following aspects: access to specialist services, queuing to wait for care, availability of appointments and schedule to schedule the appointment, Missing the appointment having arrived at the service, the priority that institutions give to maternal mothers, getting up early to get care, requesting an appointment, waiting conditions and choosing the clinic or staff. Conclusions: the previous aspects are intensified in social classes with less advantages. Reducing social inequalities is required to decrease health inequities.

[Women in pregnancy, delivery, and postpartum: a look from the feminist point of view]. / Mujeres en embarazo, parto, y posparto: una mirada desde el pensamiento feminista.

This essay analyzes the contributions of the gender approach of feminist thinking to understand the relationship of women in pregnancy, childbirth, and postpartum with health service personnel. Origi nal qualitative design research was analyzed, were the relationship of women with health personnel is visualized. It is concluded that the gender approach of feminist thinking is a useful approximation to understand the problem of women's relations with health service personnel in aspects such as obstetric violence, care and the intersection of gender with other categories, in the framework of the health system.

Este ensayo analiza los aportes del enfoque de género del pensamiento feminista para comprender la relación de las mujeres en el embarazo, parto, y posparto con el personal de los servicios de salud. Se analizaron investigaciones originales de diseño cualitativo en las cuales se visualiza la relación de las mujeres con el personal de salud. Se concluye que el enfoque de género del pensamiento feminista es una aproximación útil para comprender la problemática de las relaciones de las mujeres con el personal de los servicios de salud en aspectos como la violencia obstétrica, el cuidado y la intersección del género con otras categorías, en el marco del sistema de salud.

Relations established by women during pregnancy, delivery and postpartum with health personnel according to social class in Bogotá: qualitative study. / Relaciones que establecen las mujeres durante el embarazo, parto y posparto con el personal de salud según clase social en Bogotá: estudio cualitativo.

OBJECTIVE: To understand the relationship established between women in a situation of pregnancy, childbirth and postpartum with the health services personnel according to social class in Bogotá (Colombia). MATERIALS AND METHODS: Qualitative study. Critical hermeneutical perspective and critical ethnography. Theoretical sampling. Analysis by triangulation in Atlas.ti. 9 women and 8 health professionals participated. 38 in-depth interviews were conducted for 13 months and 62 accompaniments to the maternal in the activities of prenatal control, vaccination, labor, postpartum consultation, follow up exams, prophylactic pisco course, hospitalization and waiting room, both in public services as private. RESULTS: There are inequalities according to social class in which the relationship between women and staff is configured in the following aspects: permeability to the needs of women, recognition of psychosocial aspects, having different points of view against a medical recommendation and right to complain or demand to improve. CONCLUSIONS: The situation described above intensifies gender issues in women with a less advantageous social class. It is necessary to develop interventions in educational and health institutions that consider aspects where human resources are sensitized on social issues related to the theoretical proposals of research and the democratization of medical information. It is unfair that the condition of social class and gender affects the quality of care and economically stratifies people's rights.

OBJETIVO: Comprender la relación que se establece entre las mujeres en situación de embarazo, parto y posparto con el personal de los servicios de salud según la clase social en Bogotá (Colombia). MATERIALES Y MÉTODOS: Estudio cualitativo, perspectiva hermenéutica crítica y etnografía crítica con muestreo teórico. Análisis por triangulación en Atlas.ti. Participaron nueve mujeres y ocho profesionales de la salud. Se realizaron 38 entrevistas a profundidad durante 13 meses y 62 acompañamientos a las mujeres en las actividades de control prenatal, vacunación, trabajo de parto, consulta posparto, exámenes de seguimiento, curso psicoprofiláctico, hospitalización y sala de espera, tanto en servicios públicos como privados. RESULTADOS: Existen desigualdades según la clase social en la forma en que se configura la relación entre las mujeres y el personal en los siguientes aspectos: permeabilidad a las necesidades de la mujer, reconocimiento de aspectos psicosociales, tener diferentes puntos de vista frente a una recomendación médica y derecho a quejarse o exigir para mejorar la atención de los servicios de salud. CONCLUSIONES: Lo anterior intensifica las problemáticas relacionadas con el género en las mujeres con una clase social menos aventajada. Es necesario el desarrollo de intervenciones en las instituciones educativas y de salud que tengan en cuenta aspectos donde se sensibilice el recurso humano en temas sociales relacionados con las propuestas teóricas de la investigación y la democratización de la información médica. Es injusto que la condición de clase social y de género afecte la calidad en la atención y estratifique económicamente los derechos de las personas.

Relaciones que establecen las mujeres durante el embarazo, parto y posparto con el personal de salud según clase social en Bogotá: estudio cualitativo / Relations established by women during pregnancy, delivery and postpartum with health personnel according to social class in Bogotá: qualitative study

RESUMEN Objetivo: Comprender la relación que se establece entre las mujeres en situación de embarazo, parto y posparto con el personal de los servicios de salud según la clase social en Bogotá (Colombia). Materiales y métodos: Estudio cualitativo, perspectiva hermenéutica crítica y etnografía crítica con muestreo teórico. Análisis por triangulación en Atlas.ti. Participaron nueve mujeres y ocho profesionales de la salud. Se realizaron 38 entrevistas a profundidad durante 13 meses y 62 acompañamientos a las mujeres en las actividades de control prenatal, vacunación, trabajo de parto, consulta posparto, exámenes de seguimiento, curso psicoprofiláctico, hospitalización y sala de espera, tanto en servicios públicos como privados. Resultados: Existen desigualdades según la clase social en la forma en que se configura la relación entre las mujeres y el personal en los siguientes aspectos: permeabilidad a las necesidades de la mujer, reconocimiento de aspectos psicosociales, tener diferentes puntos de vista frente a una recomendación médica y derecho a quejarse o exigir para mejorar la atención de los servicios de salud. Conclusiones: Lo anterior intensifica las problemáticas relacionadas con el género en las mujeres con una clase social menos aventajada. Es necesario el desarrollo de intervenciones en las instituciones educativas y de salud que tengan en cuenta aspectos donde se sensibilice el recurso humano en temas sociales relacionados con las propuestas teóricas de la investigación y la democratización de la información médica. Es injusto que la condición de clase social y de género afecte la calidad en la atención y estratifique económicamente los derechos de las personas.

ABSTRACT Objective: To understand the relationship established between women in a situation of pregnancy, childbirth and postpartum with the health services personnel according to social class in Bogotá (Colombia). Materials and methods: Qualitative study. Critical hermeneutical perspective and critical ethnography. Theoretical sampling. Analysis by triangulation in Atlas.ti. 9 women and 8 health professionals participated. 38 in-depth interviews were conducted for 13 months and 62 accompaniments to the maternal in the activities of prenatal control, vaccination, labor, postpartum consultation, follow up exams, prophylactic pisco course, hospitalization and waiting room, both in public services as private. Results: There are inequalities according to social class in which the relationship between women and staff is configured in the following aspects: permeability to the needs of women, recognition of psychosocial aspects, having different points of view against a medical recommendation and right to complain or demand to improve. Conclusions: The situation described above intensifies gender issues in women with a less advantageous social class. It is necessary to develop interventions in educational and health institutions that consider aspects where human resources are sensitized on social issues related to the theoretical proposals of research and the democratization of medical information. It is unfair that the condition of social class and gender affects the quality of care and economically stratifies people's rights.

Mujeres en embarazo, parto, y posparto: una mirada desde el pensamiento feminista* / Women in pregnancy, delivery, and postpartum: a look from the feminist point of view

RESUMEN Este ensayo analiza los aportes del enfoque de género del pensamiento feminista para comprender la relación de las mujeres en el embarazo, parto, y posparto con el personal de los servicios de salud. Se analizaron investigaciones originales de diseño cualitativo en las cuales se visualiza la relación de las mujeres con el personal de salud. Se concluye que el enfoque de género del pensamiento feminista es una aproximación útil para comprender la problemática de las relaciones de las mujeres con el personal de los servicios de salud en aspectos como la violencia obstétrica, el cuidado y la intersección del género con otras categorías, en el marco del sistema de salud.

ABSTRACT This essay analyzes the contributions of the gender approach of feminist thinking to understand the relationship of women in pregnancy, childbirth, and postpartum with health service personnel. Origi nal qualitative design research was analyzed, were the relationship of women with health personnel is visualized. It is concluded that the gender approach of feminist thinking is a useful approximation to understand the problem of women's relations with health service personnel in aspects such as obstetric violence, care and the intersection of gender with other categories, in the framework of the health system.

Small-Molecule-Targeting Hairpin Loop of hTERT Promoter G-Quadruplex Induces Cancer Cell Death.

Increased telomerase activity is associated with malignancy and poor prognosis in human cancer, but the development of targeted agents has not yet provided clinical benefit. Here we report that, instead of targeting the telomerase enzyme directly, small molecules that bind to the G-hairpin of the hTERT G-quadruplex-forming sequence kill selectively malignant cells without altering the function of normal cells. RG260 targets the hTERT G-quadruplex stem-loop folding but not tetrad DNAs, leading to downregulation of hTERT expression. To improve physicochemical and pharmacokinetic properties, we derived a small-molecule analog, RG1603, from the parent compound. RG1603 induces mitochondrial defects including PGC1α and NRF2 inhibition and increases oxidative stress, followed by DNA damage and apoptosis. RG1603 injected as a single agent has tolerable toxicity while achieving strong anticancer efficacy in a tumor xenograft mouse model. These results demonstrate a unique approach to inhibiting the hTERT that functions by impairing mitochondrial activity, inducing cell death.

Targeting PIM Kinase with PD1 Inhibition Improves Immunotherapeutic Antitumor T-cell Response.

PURPOSE: Adoptive T-cell therapy (ACT) of cancer, which involves the infusion of ex vivo-engineered tumor epitope reactive autologous T cells into the tumor-bearing host, is a potential treatment modality for cancer. However, the durable antitumor response following ACT is hampered either by loss of effector function or survival of the antitumor T cells. Therefore, strategies to improve the persistence and sustain the effector function of the antitumor T cells are of immense importance. Given the role of metabolism in determining the therapeutic efficacy of T cells, we hypothesize that inhibition of PIM kinases, a family of serine/threonine kinase that promote cell-cycle transition, cell growth, and regulate mTORC1 activity, can improve the potency of T cells in controlling tumor. EXPERIMENTAL DESIGN: The role of PIM kinases in T cells was studied either by genetic ablation (PIM1-/-PIM2-/-PIM3-/-) or its pharmacologic inhibition (pan-PIM kinase inhibitor, PimKi). Murine melanoma B16 was established subcutaneously and treated by transferring tumor epitope gp100-reactive T cells along with treatment regimen that involved inhibiting PIM kinases, anti-PD1 or both. RESULTS: With inhibition of PIM kinases, T cells had significant reduction in their uptake of glucose, and upregulated expression of memory-associated genes that inversely correlate with glycolysis. In addition, the expression of CD38, which negatively regulates the metabolic fitness of the T cells, was also reduced in PimKi-treated cells. Importantly, the efficacy of antitumor T-cell therapy was markedly improved by inhibiting PIM kinases in tumor-bearing mice receiving ACT, and further enhanced by adding anti-PD1 antibody to this combination. CONCLUSIONS: This study highlights the potential therapeutic significance of combinatorial strategies where ACT and inhibition of signaling kinase with checkpoint blockade could improve tumor control.

Returning to work after breast cancer: A systematic review of reviews.

BACKGROUND: Due to improvements in breast cancer diagnosis and treatment, the healthcare system faces a growing number of cancer survivors. Breast cancer survivors experience many difficulties when returning to work, including discrimination at work and lack of support by employers and colleagues. OBJECTIVE: To point out the knowledge in literature up to date about return to work (RTW) after breast cancer, the factors influencing it and the interventions to facilitate it. METHODS: A literature search was conducted in January 2017 using the databases Medline (PubMed) and Scopus. Studies were included if they analyzed the problem of RTW in women treated for breast cancer. RESULTS: Twenty-six articles met the inclusion criteria. The studies were divided into four themes: factors facilitating or impeding RTW; interventions to enhance RTW; lived experiences of RTW; economic aspects related to cancer survivors and RTW. CONCLUSIONS: The heterogeneity of the interventions suggests the need for a better definition of the concept of RTW. To compare interventions, studies should use a rigorous approach and better outcome measures should be identified to evaluate RTW.

Mechanisms Behind Resistance to PI3K Inhibitor Treatment Induced by the PIM Kinase.

Cancer resistance to PI3K inhibitor therapy can be in part mediated by increases in the PIM1 kinase. However, the exact mechanism by which PIM kinase promotes tumor cell resistance is unknown. Our study unveils the pivotal control of redox signaling by PIM kinases as a driver of this resistance mechanism. PIM1 kinase functions to decrease cellular ROS levels by enhancing nuclear factor erythroid 2-related factor 2 (NRF2)/antioxidant response element activity. PIM prevents cell death induced by PI3K-AKT-inhibitory drugs through a noncanonical mechanism of NRF2 ubiquitination and degradation and translational control of NRF2 protein levels through modulation of eIF4B and mTORC1 activity. Importantly, PIM also controls NAD(P)H production by increasing glucose flux through the pentose phosphate shunt decreasing ROS production, and thereby diminishing the cytotoxicity of PI3K-AKT inhibitors. Treatment with PIM kinase inhibitors reverses this resistance phenotype, making tumors increasingly susceptible to small-molecule therapeutics, which block the PI3K-AKT pathway.

Paget's disease of the male breast: case report and a point of view from actual literature.

INTRODUCTION: Paget disease of the nipple in man is a very rare breast cancer, and there are not standard procedures or guidelines. In any cases, a Paget's disease could hide an invasive ductal breast cancer. CASE DESCRIPTION: We report the case of a 77-years old man affected by Alzheimer's disease, who presented to our attention because of an ulcerated palpable mass in the right nipple. A biopsy of the lesion showed "intra-epidermic proliferation of epitelioid cells, associated with linfo-plasmacellular infiltration of superficial dermis, compatible with Paget's disease (pTis)". We discussed the case in the multidisciplinary meeting and decided to subject the patient to surgery, so a right mastectomy plus sentinel lymph node biopsy (SLNB) were performed. Histo-pathological examination revealed "invasive ductal carcinoma of the breast, associated with a small component of in situ ductal carcinoma and Paget's disease of the nipple with superficial ulceration". Resection margins were free. Sentinel lymph node was negative. Biological features were as follows: ER 95%, PR 60%, Her-2/neu 1+, Ki-67 35%. The patient was discharged in the third post-operative day in good conditions. In the following weeks the patient's healing process was good and free of complications. CONCLUSIONS: Clinical recognition of Paget's disease is very important also in man, because it can be the alarm bell for an underlying invasive ductal breast cancer, often more aggressive than in woman.

Targeting the PIM protein kinases for the treatment of a T-cell acute lymphoblastic leukemia subset.

New approaches are needed for the treatment of patients with T-cell acute lymphoblastic leukemia (T-ALL) who fail to achieve remission with chemotherapy. Analysis of the effects of pan-PIM protein kinase inhibitors on human T-ALL cell lines demonstrated that the sensitive cell lines expressed higher PIM1 protein kinase levels, whereas T-ALL cell lines with NOTCH mutations tended to have lower levels of PIM1 kinase and were insensitive to these inhibitors. NOTCH-mutant cells selected for resistance to gamma secretase inhibitors developed elevated PIM1 kinase levels and increased sensitivity to PIM inhibitors. Gene profiling using a publically available T-ALL dataset demonstrated overexpression of PIM1 in the majority of early T-cell precursor (ETP)-ALLs and a small subset of non-ETP ALL. While the PIM inhibitors blocked growth, they also stimulated ERK and STAT5 phosphorylation, demonstrating that activation of additional signaling pathways occurs with PIM inhibitor treatment. To block these pathways, Ponatinib, a broadly active tyrosine kinase inhibitor (TKI) used to treat chronic myelogenous leukemia, was added to this PIM-inhibitor regimen. The combination of Ponatinib with a PIM inhibitor resulted in synergistic T-ALL growth inhibition and marked apoptotic cell death. Treatment of mice engrafted with human T-ALL with these two agents significantly decreased the tumor burden and improved the survival of treated mice. This dual therapy has the potential to be developed as a novel approach to treat T-ALL with high PIM expression.

Insulin receptor substrate 1 is a substrate of the Pim protein kinases.

The Pim family of serine/threonine protein kinases (Pim 1, 2, and 3) contribute to cellular transformation by regulating glucose metabolism, protein synthesis, and mitochondrial oxidative phosphorylation. Drugs targeting the Pim protein kinases are being tested in phase I/II clinical trials for the treatment of hematopoietic malignancies. The goal of these studies was to identify Pim substrate(s) that could help define the pathway regulated by these enzymes and potentially serve as a biomarker of Pim activity. To identify novel substrates, bioinformatics analysis was carried out to identify proteins containing a consensus Pim phosphorylation site. This analysis identified the insulin receptor substrate 1 and 2 (IRS1/2) as potential Pim substrates. Experiments were carried out in tissue culture, animals, and human samples from phase I trials to validate this observation and define the biologic readout of this phosphorylation. Our study demonstrates in both malignant and normal cells using either genetic or pharmacological inhibition of the Pim kinases or overexpression of this family of enzymes that human IRS1S1101 and IRS2S1149 are Pim substrates. In xenograft tumor experiments and in a human phase I clinical trial, a pan-Pim inhibitor administered in vivo to animals or humans decreased IRS1S1101 phosphorylation in tumor tissues. This phosphorylation was shown to have effects on the half-life of the IRS family of proteins, suggesting a role in insulin or IGF signaling. These results demonstrate that IRS1S1101 is a novel substrate for the Pim kinases and provide a novel marker for evaluation of Pim inhibitor therapy.

Aroma volatiles obtained at harvest by HS-SPME/GC-MS and INDEX/MS-E-nose fingerprint discriminate climacteric behaviour in melon fruit.

BACKGROUND: Melon aroma volatiles were extracted at harvest from juice of a climacteric near-isogenic line (NIL) SC3-5-1 with two quantitative trait loci (QTLs) introgressed which produced climacteric behaviour and its non-climacteric parental (PS) using two methodologies of analysis: static headspace solid phase micro-extraction (HS-SPME) by gas chromatography-mass spectrometry (GC-MS) and inside needle dynamic extraction (INDEX) by MS-based electronic nose (MS-E-nose). RESULTS: Of the 137 volatiles compounds identified, most were found at significantly higher concentrations in SC3-5-1 than in PS in both seasons. These volatiles were mostly esters, alcohols, sulfur-derived esters and even some aldehydes and others. The number of variables with high correlation values was reduced by using correlation network analysis. Partial least squares-discriminant analysis (PLS-DA) achieved the correct classification of PS and SC3-5-1. The ions m/z 74, 91, 104, 105, 106 and 108, mainly volatile derivatives precursor phenylalanine, were the most discriminant in SC3-5-1 and PS. As many as 104 QTLs were mapped in season 1 and at least 78 QTLs in each season with an effect above the PS mean. CONCLUSION: GC-MS gave better discrimination than E-nose. Most of the QTLs that mapped in both seasons enhanced aroma volatiles associated with climacteric behaviour.

Hydroxytyrosol and potential uses in cardiovascular diseases, cancer, and AIDS.

Hydroxytyrosol is one of the main phenolic components of olive oil. It is present in the fruit and leaf of the olive (Olea europaea L.). During the past decades, it has been well documented that this phenolic compound has health benefits and a protective action has been found in preclinical studies against several diseases. Here, we review its bioavailability in human beings and several assays showing significant results related with cardiovascular diseases, cancer, and acquired immunodeficiency syndrome (AIDS). Mechanisms of action include potent anti-oxidant and anti-inflammatory effects, among others. The importance of hydroxytyrosol in protection of low-density lipoproteins and consequently its implication in the reduction of cardiovascular disease risk has been highlighted by the European Food Safety Authority, concluding that 5 mg of hydroxytyrosol and its derivatives should be consumed daily to reach this effect at physiological level. We discuss the potential uses of this compound in supplements, nutraceutic foods, or topical formulations in the disease risk reduction. Finally, we conclude that more studies are needed to sustain or reject many other health claims not yet fully documented and to validate these newly available hydroxytyrosol-based products, because it seems to be a good candidate to reduce the risk of diseases mentioned.

Leptin and fasting regulate rat gastric glucose-regulated protein 58.

The stomach secretes a wide range of peptides with essential metabolic functions, and thereby plays an important role in the regulation of energy homeostasis. Disulfide isomerase glucose-regulated protein 58 (GRp58) is a molecular chaperone member of the endoplasmic reticulum (ER) stress signaling pathway, which is a marker for human gastric cancer. Since GRp58 seems to be regulated by a phosphorylation/dephosphorylation pattern shift, we used the 2DE gel methodology and peptide mass fingerprinting-protein identification by means of MALDI-TOF mass spectrometry. We show that gastric mucosa GRp58 is dephosphorylated by fasting, and this effect is blunted when fasted rats are treated with leptin. Furthermore, we assessed the gene expression of GRp58 under different physiological settings known to be associated with energy homeostasis (fasting, leptin treatment and leptin deficiency). We found that intraperitoneal administration of leptin increases whereas leptin deficiency decreases GRp58 mRNA levels. However, GRp58 expression remains unchanged after fasting, indicating that leptin actions on GRp58 are no direct sensitivity to fasting. Dissection of the molecular pathways mediating the interactions between ER stress-related factors and nutrient availability, as well as their target genes, may open a new avenue for the study of obesity and other metabolic disorders.

Nonnutritive sucking habits among preschool-aged children.

OBJECTIVE: To verify the prevalence and factors associated with nonnutritive sucking habits in preschool-aged children attending daycare centers and preschools in Natal, Brazil. METHODS: A cross-sectional study was conducted with 1,190 children of both sexes, aged 3 to 5 years, enrolled in daycare centers and preschools in Natal, Brazil. Exclusion criteria were: children with cleft lip and palate, temporomandibular joint disorders, or submitted to orthodontic and/or orthopedic treatment; as well as institutions specialized in children with disabilities. Parents or guardians answered a structured questionnaire providing information on the institution, children's sex and age, parents' educational level, and habit-related questions. Data analysis was performed using the chi-square test and logistic regression. RESULTS: A prevalence of 40.2% of nonnutritive sucking habits was obtained; of these, 27.7% were pacifier-sucking and 12.5% were finger-sucking habits. Girls showed a higher percentage of sucking habits, especially finger sucking (p = 0.02); younger children showed a higher prevalence of pacifier-sucking habits (p = 0.0006). A higher frequency of pacifier- and finger-sucking habits was associated, respectively, with parents' higher education (p < 0.05) and elementary education (p < 0.05). Logistic regression revealed that younger individuals (p = 0.033) and secondary education level of parents (p = 0.035) are independent factors for habit persistence. CONCLUSION: A high prevalence of nonnutritive sucking habits was verified, highlighting younger age groups and secondary education level of parents as important associated factors.