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Las enfermedades de la tiroides, tanto benignas como malignas, son altamente prevalentes a nivel mundial, por lo que es muy probable que durante la pandemia por SARS-CoV-2 veamos pacientes con ambas enfermedades. Esto exige el conocimiento de las implicaciones potenciales de este nuevo virus en el funcionamiento de la glándula, en los tratamientos usuales para estas enfermedades y en consideraciones especiales para este grupo poblacional. A la fecha no hay evidencia que soporte que las enfermedades tiroideas aumenten el riesgo de infección o severidad de la enfermedad. Sin embargo, es posible que durante infecciones severas por SARS-CoV-2 en personas con o sin antecedente de enfermedad tiroidea puedan presentar alteración de las pruebas tiroideas, aunque transitoriamente y sin requerimiento de tratamiento específico. Es fundamental que los pacientes continúen con sus tratamientos ambulatorios y se difiera, en la medida de lo posible, los procedimientos quirúrgicos o la administración de yodo radioactivo hasta que se considere seguro realizarlos.

Desde finales del año 2019 un nuevo coronavirus, SARS-CoV-2, se extendió desde China hacia el resto del mundo, causando la pandemia de una enfermedad denominada COVID-19. Una enfermedad sistémica que en algunos casos produce neumonía severa que incluso puede progresar a falla respiratoria aguda y finalmente la muerte. Entre las comorbilidades que se han asociado con un aumento en la mortalidad por SARS-CoV-2 se encuentra la diabetes. En general, se estima que tener diabetes aumenta un 18 % el riesgo de infecciones respiratorias, en parte por el impacto que genera sobre la inmunidad innata o adquirida, lo que estaría contribuyendo a una presentación clínica más severa del SARS-CoV-2 al comparar con población sin diabetes. Considerando que existe una asociación entre mal control glucémico y mayor severidad clínica de la infección por COVID-19, se deben hacer importantes consideraciones sobre el manejo farmacológico brindado a los pacientes; el perfilamiento dependerá de las condiciones de cada paciente, de la severidad de la enfermedad y del tipo de manejo instaurado ya sea ambulatorio o intrahospitalario.

Objective This study aimed to estimate the effectiveness of a comprehensive diabetes program (CDP) in terms of glycemic control, adherence, and the selection of candidates for sensor-augmented insulin pump therapy (SAP). Methods We compared diabetes control before and 6 months after CDP. The program was based on disease management using a logical model dealing with the following: case management, education and coaching, nutritional assessment, and mental health. Results The CDP improved glycemic control, HbA1c decreased by 0.56% (p-value = 0.004; 95% CI: 0.14–0.98) and 19.1% of the patients reached the HbA1c goal without hypoglycemia. The CDP reduced by 52.4% the indication for SAP due to better glycemic control (36.4%) or non-adherence issues (63.6%); the remaining 47.6% persisted with poor glycemic control despite good adherence and were scaled to SAP. Among the 30 suitable candidates for SAP therapy, 60% did not reach the HbA1c goal and 40% had either hypoglycemic episodes (severe or persistent) or dawn phenomenon. The overall non-adherence rate was 33.3%. Conclusions CDP optimized the selection of suitable candidates for SAP by improving glycemic control and identifying adherence issues early. These results provide evidence of the impact of the implementation of patient selection and educational protocols in the real-life setting of a highly experienced clinic.

To evaluate the effectiveness and safety of sensor-augmented insulin pump therapy (SAP) in addition to a comprehensive diabetes program on glycated hemoglobin (HbA1c), severe hypoglycemia, ketoacidosis, and the hospital admission rate in patients with type 1 diabetes under real-world settings during a 2-year follow-up.This was a retrospective real-life study comparing diabetes control before and after SAP therapy initiation. Patients ≥18 years old with type 1 diabetes were included. They were followed for 2 years with clinical assessments at months 3, 6, 12, 18, and 24. Effectiveness was estimated by difference in medians of HbA1c from baseline and at each follow-up visit. Safety was assessed by comparing the annual rates of severe hypoglycemia, hyperglycemic crisis, and hospital admission related to diabetes.162 patients were included, median age 32 years, women 73%). The main indication for SAP was poor metabolic control (51.2%). At 2 years HbA1c decreased from 8.4% to 7.5% (-0.9%, 95% CI: 0.5-1.2; p0.0001), HbA1c ≤7% improved from 14.2% to 25.3% (11.1%, 95% CI: 19.7-2.5; p=0.006), and severe hypoglycemia decreased from 22.2% to 14.1% (-8.1%, 95% CI: -16.5 to 0.3; p=0.03).SAP therapy improved glycemic control after the third month of use and for up to 2 years of follow-up, with lower rates of hospital admission and severe hypoglycemia. More studies are needed to assess the add-on impact of education programs and technologies for diabetes care.

// Victor M. Blanco 1 , Zhengtao Chu 1, 2 , Kathleen LaSance 3 , Brian D. Gray 4 , Koon Yan Pak 4 , Therese Rider 5 , Kenneth D. Greis 5 , Xiaoyang Qi 1, 2 1 Division of Hematology-Oncology, Department of Internal Medicine, University of Cincinnati College of Medicine, Cincinnati, Ohio 45267, USA 2 Division of Human Genetics, Department of Pediatrics, Cincinnati Children’s Hospital Medical Center, Cincinnati, Ohio 45229, USA 3 Department of Radiology, University of Cincinnati College of Medicine, Cincinnati, Ohio 45267, USA 4 Molecular Targeting Technologies, Inc., West Chester, Pennsylvania 19380, USA 5 Department of Cancer Biology, University of Cincinnati College of Medicine, Cincinnati, Ohio 45267, USA Correspondence to: Xiaoyang Qi, email: xiaoyang.qi@uc.edu Keywords: glioblastoma, liposome, PET, optical imaging, SapC-DOPS Abbreviations: SapC, Saposin C; DOPS, dioleylphosphatidylserine; PS, phosphatidylserine Received: June 25, 2015 Accepted: March 28, 2016 Published: April 16, 2016 ABSTRACT Multimodal tumor imaging with targeted nanoparticles potentially offers both enhanced specificity and sensitivity, leading to more precise cancer diagnosis and monitoring. We describe the synthesis and characterization of phenol-substituted, lipophilic orange and far-red fluorescent dyes and a simple radioiodination procedure to generate a dual (optical and nuclear) imaging probe. MALDI-ToF analyses revealed high iodination efficiency of the lipophilic reporters, achieved by electrophilic aromatic substitution using the chloramide 1,3,4,6-tetrachloro-3α,6α-diphenyl glycoluril (Iodogen) as the oxidizing agent in an organic/aqueous co-solvent mixture. Upon conjugation of iodine-127 or iodine-124-labeled reporters to tumor-targeting SapC-DOPS nanovesicles, optical (fluorescent) and PET imaging was performed in mice bearing intracranial glioblastomas. In addition, tumor vs non-tumor (normal brain) uptake was compared using iodine-125. These data provide proof-of-principle for the potential value of SapC-DOPS for multimodal imaging of glioblastoma, the most aggressive primary brain tumor.

Background— High-fat diet (HFD) promotes endothelial dysfunction and proinflammatory monocyte activation, which contribute to atherosclerosis in obesity. We investigated whether HFD also induces the dysfunction of red blood cells (RBCs), which serve as a reservoir for chemokines via binding to Duffy antigen receptor for chemokines (DARC). Methods and Results— A 60% HFD for 12 weeks, which produced only minor changes in lipid profile in C57/BL6 mice, markedly augmented the levels of monocyte chemoattractant protein-1 bound to RBCs, which in turn stimulated macrophage migration through an endothelial monolayer. Levels of RBC-bound KC were also increased by HFD. These effects of HFD were abolished in DARC –/– mice. In RBCs from HFD-fed wild-type and DARC –/– mice, levels of membrane cholesterol and phosphatidylserine externalization were increased, fostering RBC-macrophage inflammatory interactions and promoting macrophage phagocytosis in vitro. When labeled ex vivo and injected into wild-type mice, RBCs from HFD-fed mice exhibited ≈3-fold increase in splenic uptake. Finally, RBCs from HFD-fed mice induced increased macrophage adhesion to the endothelium when they were incubated with isolated aortic segments, indicating endothelial activation. Conclusions— RBC dysfunction, analogous to endothelial dysfunction, occurs early during diet-induced obesity and may serve as a mediator of atherosclerosis. These findings may have implications for the pathogenesis of atherosclerosis in obesity, a worldwide epidemic.

Pancreatic cancer remains one of the most intractable cancers, with a dismal prognosis reflected by a 5-year survival of ~6%. Since early disease symptoms are undefined and specific biomarkers are lacking, about 80% of patients present with advanced, inoperable tumors that represent a daunting challenge. Despite many clinical trials, no single chemotherapy agent has been reliably associated with objective response rates above 10% or median survival longer than 5 to 7 months. Although combination chemotherapy regimens have in recent years provided some improvement, overall survival (8-11 months) remains very poor. There is therefore a critical need for novel therapies that can improve outcomes for pancreatic cancer patients. Here, we present a summary of the current therapies used in the management of advanced pancreatic cancer and review novel therapeutic strategies that target tumor biomarkers. We also describe our recent research using phosphatidylserine-targeted saposin C–coupled dioleoylphosphatidylserine nanovesicles for imaging and therapy of pancreatic cancer.

Glioblastoma multiforme (GBM) is the most common malignant primary brain tumor. Classified by the World Health Organization (WHO) as grade IV astrocytoma, GBMs are extremely aggressive, almost always recur, and despite our best efforts, remain incurable. This review describes the traditional treatment approaches that led to moderate successes in GBM patients, discusses standard imaging modalities, and presents data supporting the use of SapC-DOPS, a novel proteoliposomal formulation with tumoricidal activity, as a promising diagnostic imaging tool and an innovative anti-cancer agent against GBM.

Oxidative stress plays a fundamental role in abdominal aortic aneurysm (AAA) formation. Activated polymorphonuclear leukocytes (or neutrophils) are associated with AAA and express myeloperoxidase (MPO), which promotes inflammation, matrix degradation, and other pathological features of AAA, including enhanced oxidative stress through generation of reactive oxygen species. Both plasma and aortic MPO levels are elevated in patients with AAA, but the role of MPO in AAA pathogenesis has, heretofore, never been investigated. Here, we show that MPO gene deletion attenuates AAA formation in two animal models: ANG II infusion in apolipoprotein E-deficient mice and elastase perfusion in C57BL/6 mice. Oral administration of taurine [1% or 4% (wt/vol) in drinking water], an amino acid known to react rapidly with MPO-generated oxidants like hypochlorous acid, also prevented AAA formation in the ANG II and elastase models as well as the CaCl2 application model of AAA formation while reducing aortic peroxidase activity and aortic protein-bound dityrosine levels, an oxidative cross link formed by MPO. Both MPO gene deletion and taurine supplementation blunted aortic macrophage accumulation, elastin fragmentation, and matrix metalloproteinase activation, key features of AAA pathogenesis. Moreover, MPO gene deletion and taurine administration significantly attenuated the induction of serum amyloid A, which promotes ANG II-induced AAAs. These data implicate MPO in AAA pathogenesis and suggest that studies exploring whether taurine can serve as a potential therapeutic for the prevention or treatment of AAA in patients merit consideration. NEW & NOTEWORTHY Neutrophils are abundant in abdominal aortic aneurysm (AAA), and myeloperoxidase (MPO), prominently expressed in neutrophils, is associated with AAA in humans. This study demonstrates that MPO gene deletion or supplementation with the natural product taurine, which can scavenge MPO-generated oxidants, can prevent AAA formation, suggesting an attractive potential therapeutic strategy for AAA.

// Victor M. Blanco 1 , Zhengtao Chu 1,2 , Subrahmanya D. Vallabhapurapu 1 , Mahaboob K. Sulaiman 1 , Ady Kendler 3 , Olivier Rixe 4 , Ronald E. Warnick 5 , Robert S. Franco 1 and Xiaoyang Qi 1,2 1 Division of Hematology and Oncology, Department of Internal Medicine, University of Cincinnati College of Medicine, Cincinnati, Ohio 2 Division of Human Genetics, Department of Pediatrics, Cincinnati Children’s Hospital Medical Center, Cincinnati, Ohio 3 Department of Pathology and Laboratory Medicine, University of Cincinnati College of Medicine, Cincinnati, Ohio 4 Division of Hematology/Oncology, Georgia Regents University, GRU Cancer Center, Augusta, Georgia 5 Department of Neurosurgery, University of Cincinnati Brain Tumor Center, and Mayfield Clinic, Cincinnati, Ohio Correspondence: Xiaoyang Qi, email: // Keywords : Glioblastoma; brain metastasis; SapC-DOPS; imaging; cancer therapy Received : June 2, 2014 Accepted: July 13, 2014 Published: July 14, 2014 Abstract Brain tumors, either primary (e.g., glioblastoma multiforme) or secondary (metastatic), remain among the most intractable and fatal of all cancers. We have shown that nanovesicles consisting of Saposin C (SapC) and dioleylphosphatidylserine (DOPS) are able to effectively target and kill cancer cells both in vitro and in vivo . These actions are a consequence of the affinity of SapC-DOPS for phosphatidylserine, an acidic phospholipid abundantly present in the outer membrane of a variety of tumor cells and tumor-associated vasculature. In this study, we first characterize SapC-DOPS bioavailability and antitumor effects on human glioblastoma xenografts, and confirm SapC-DOPS specificity towards phosphatidylserine by showing that glioblastoma targeting is abrogated after in vivo exposure to lactadherin, which binds phosphatidylserine with high affinity. Second, we demonstrate that SapC-DOPS selectively targets brain metastases-forming cancer cells both in vitro , in co-cultures with human astrocytes, and in vivo , in mouse models of brain metastases derived from human breast or lung cancer cells. Third, we demonstrate that SapC-DOPS nanovesicles have cytotoxic activity against metastatic breast cancer cells in vitro , and prolong the survival of mice harboring brain metastases. Taken together, these results support the potential of SapC-DOPS for the diagnosis and therapy of primary and metastatic brain tumors.

During chronic caloric excess, adipose tissue expands primarily by enlargement of individual adipocytes, which become stressed with lipid overloading, thereby contributing to obesity-related disease. Although adipose tissue contains numerous preadipocytes, differentiation into functionally competent adipocytes is insufficient to accommodate the chronic caloric excess and prevent adipocyte overloading. We report for the first time that a chronic high-fat diet (HFD) impairs adipogenic differentiation, leading to accumulation of inefficiently differentiated adipocytes with blunted expression of adipogenic differentiation-specific genes. Preadipocytes from these mice likewise exhibit impaired adipogenic differentiation, and this phenotype persists during in vitro cell culture. HFD-induced impaired adipogenic differentiation is associated with elevated expression of histone deacetylase 9 (HDAC9), an endogenous negative regulator of adipogenic differentiation. Genetic ablation of HDAC9 improves adipogenic differentiation and systemic metabolic state during an HFD, resulting in diminished weight gain, improved glucose tolerance and insulin sensitivity, and reduced hepatosteatosis. Moreover, compared with wild-type mice, HDAC9 knockout mice exhibit upregulated expression of beige adipocyte marker genes, particularly during an HFD, in association with increased energy expenditure and adaptive thermogenesis. These results suggest that targeting HDAC9 may be an effective strategy for combating obesity-related metabolic disease.

This paper describes various reliability concerns of the newly developed INGRID detector. This radiation detector is fabricated by waferscale CMOS post-processing; fresh detectors show excellent performance. Since the microsystems will be used unpackaged they are susceptible to all kinds of environmental conditions. The device passed tests of micro-ESD, radiation hardness, dielectric strength; but humidity tests show one weakness of SU-8 as a structural material. Already after 1 day of exposure to a humid condition the structural integrity, as measured by a shear stress test, is dramatically lowered. Dry storage of these devices is therefore a necessity. KMPR photoresist shows promising results as an alternative structural material.

Normal brain function requires proper supply of oxygen and glucose in a timely and local manner. This is achieved through an orchestrated intercellular communication between neurones, astrocytes and microvessels that results in a rapid and restricted increase in cerebral blood flow, a process known as neurovascular coupling. Astrocytic end-feet make close contacts with neuronal synapses and blood vessels and, given their ability to release vasoactive signals following neuronal activation, have been recognized as key intermediaries in the neurovascular response. Both dilating and constricting signals appear to be released from astrocytes upon increases in intracellular Ca(2+) concentration, and both dilatation and constriction of brain vessels have been observed in previous studies. In this article, we discuss the various astrocyte-derived vasodilating and vasoconstricting signals, their interactions and effects on astrocytes and vascular smooth muscle cells, and suggest the importance of the intrinsic properties of the latter cell type on the overall neurovascular response. We present a working model in which the rise in astrocytic Ca(2+) following neuronal activation leads not only to the rapid activation of calcium-activated K(+) channels in astrocytic end-feet, but also to their modulation by metabolites of the arachidonic acid pathway, which in general have been proposed to act on vascular smooth muscle cells rather than on astrocytes. We propose that this latter mechanism may in turn modulate K(+) signalling from astrocytes to smooth muscle cells, influencing the overall effects of the vasodilating and vasoconstricting signals released during neuronal activation.

Basal and activity-dependent cerebral blood flow changes are coordinated by the action of critical processes, including cerebral autoregulation, endothelial-mediated signaling, and neurovascular coupling. The goal of our study was to determine whether astrocytes contribute to the regulation of parenchymal arteriole (PA) tone in response to hemodynamic stimuli (pressure/flow). Cortical PA vascular responses and astrocytic Ca2+dynamics were measured using anin vitrorat/mouse brain slice model of perfused/pressurized PAs; studies were supplemented within vivoastrocytic Ca2+imaging.In vitro, astrocytes responded to PA flow/pressure increases with an increase in intracellular Ca2+. Astrocytic Ca2+responses were corroboratedin vivo, where acute systemic phenylephrine-induced increases in blood pressure evoked a significant increase in astrocytic Ca2+.In vitro, flow/pressure-evoked vasoconstriction was blunted when the astrocytic syncytium was loaded with BAPTA (chelating intracellular Ca2+) and enhanced when high Ca2+or ATP were introduced to the astrocytic syncytium. Bath application of either the TRPV4 channel blocker HC067047 or purinergic receptor antagonist suramin blunted flow/pressure-evoked vasoconstriction, whereas K+and 20-HETE signaling blockade showed no effect. Importantly, we found TRPV4 channel expression to be restricted to astrocytes and not the endothelium of PA. We present evidence for a novel role of astrocytes in PA flow/pressure-evoked vasoconstriction. Our data suggest that astrocytic TRPV4 channels are key molecular sensors of hemodynamic stimuli and that a purinergic, glial-derived signal contributes to flow/pressure-induced adjustments in PA tone. Together our results support bidirectional signaling within the neurovascular unit and astrocytes as key modulators of PA tone.

Background High toxicity, morbidity and secondary malignancy render chemotherapy of neuroblastoma inefficient, prompting the search for novel compounds. Nanovesicles offer great promise in imaging and treatment of cancer. SapC-DOPS, a stable nanovesicle formed from the lysosomal protein saposin C and dioleoylphosphatidylserine possess strong affinity for abundantly exposed surface phosphatidylserine on cancer cells. Here, we show that SapC-DOPS effectively targets and suppresses neuroblastoma growth and elucidate the molecular mechanism of SapC-DOPS action in neuroblastoma in vitro. Methods In vivo targeting of neuroblastoma was assessed in xenograft mice injected intravenously with fluorescently-labeled SapC-DOPS. Xenografted tumors were also used to demonstrate its therapeutic efficacy. Apoptosis induction in vivo was evaluated in tumor sections using the TUNEL assay. The mechanisms underlying the induction of apoptosis by SapC-DOPS were addressed through measurements of cell viability, mitochondrial membrane potential (ΔΨM), flow cytometric DNA fragmentation assays and by immunoblot analysis of second mitochondria-derived activator of caspases (Smac), Bax, Cytochrome c (Cyto c) and Caspase-3 in the cytosol or in mitochondrial fractions of cultured neuroblastoma cells. Results SapC-DOPS showed specific targeting and prevented the growth of human neuroblastoma xenografts in mice. In neuroblastoma cells in vitro, apoptosis occurred via a series of steps that included: (1) loss of ΔΨM and increased mitochondrial superoxide formation; (2) cytosolic release of Smac, Cyto c, AIF; and (3) mitochondrial translocation and polymerization of Bax. ShRNA-mediated Smac knockdown and V5 peptide-mediated Bax inhibition decreased cytosolic Smac and Cyto c release along with caspase activation and abrogated apoptosis, indicating that Smac and Bax are critical mediators of SapC-DOPS action. Similarly, pretreatment with the mitochondria-stabilizing agent bongkrekic acid decreased apoptosis indicating that loss of ΔΨM is critical for SapC-DOPS activity. Apoptosis induction was not critically dependent on reactive oxygen species (ROS) production and Cyclophilin D, since pretreatment with N-acetyl cysteine and cyclosporine A, respectively, did not prevent Smac or Cyto c release. Conclusions Taken together, our results indicate that SapC-DOPS acts through a mitochondria-mediated pathway accompanied by an early release of Smac and Bax. Specific tumor-targeting capacity and anticancer efficacy of SapC-DOPS supports its potential as a dual imaging and therapeutic agent in neuroblastoma therapy. Electronic supplementary material The online version of this article (doi:10.1186/s12943-015-0336-y) contains supplementary material, which is available to authorized users.

Lung cancer is the deadliest type of cancer for both men and women. In this study, we evaluate the in vitro and in vivo efficacy of a biotherapeutic agent composed of a lysosomal protein (Saposin C, SapC) and a phospholipid (dioleoylphosphatidylserine, DOPS), which can be assembled into nanovesicles (SapC–DOPS) with selective antitumor activity. SapC–DOPS targets phosphatidylserine, an anionic phospholipid preferentially exposed in the surface of cancer cells and tumor-associated vasculature. Because binding of SapC to phosphatidylserine is favored at acidic pHs, and the latter characterizes the milieu of many solid tumors, we tested the effect of pH on the binding capacity of SapC–DOPS to lung tumor cells. Results showed that SapC–DOPS binding to cancer cells was more pronounced at low pH. Viability assays on a panel of human lung tumor cells showed that SapC–DOPS cytotoxicity was positively correlated with cell surface phosphatidylserine levels, whereas mitochondrial membrane potential measurements were consistent with apoptosis-related cell death. Using a fluorescence tracking method in live mice, we show that SapC–DOPS specifically targets human lung cancer xenografts, and that systemic therapy with SapC–DOPS induces tumor apoptosis and significantly inhibits tumor growth. These results suggest that SapC–DOPS nanovesicles are a promising treatment option for lung cancer. Mol Cancer Ther; 14(2); 491–8. ©2015 AACR.

Regulation and maintenance of cell water volume and intracellular pH (pH i ) are vital functions that are inter-dependent; cell volume regulation affects, and is in turn affected by, changes in pH i . Disruption of either function underliesvarious pathologies. To study the interaction and kinetics of these two mechanisms, we developed and validated a quantitativefluorescence imaging microscopy method to measure simultaneous changes in pH i and volume in single cells loaded with thefluorescent probe BCECF. CWV is measured at the excitation isosbestic wavelength, whereas pH i is determined ratiometrically.The method has a time resolution of ,1 s and sensitivity to osmotic changes of ;1%. It can be applied in real time to virtuallyany cell type attached to a coverslip, independently of cellular shape and geometry. Calibration procedures and algorithms devel-oped to transform fluorescence signals into changes in cell water volume (CWV) and examples of applications are presented. INTRODUCTIONMechanisms controlling cellular pH and water volume areamong the evolutionarily oldest homeostatic functions. Pre-sumably, these two vital mechanisms appeared when macro-molecules were encased within a semipermeable plasmamembrane creating an internal compartment in which meta-bolic reactions occurred (1,2). Mechanisms controlling cellwater volume and intracellular pH (pH

We describe a multi-angle rotational optical imaging (MAROI) system for in vivo monitoring of physiopathological processes labeled with a fluorescent marker. Mouse models (brain tumor and arthritis) were used to evaluate the usefulness of this method. Saposin C (SapC)-dioleoylphosphatidylserine (DOPS) nanovesicles tagged with CellVue Maroon (CVM) fluorophore were administered intravenously. Animals were then placed in the rotational holder (MARS) of the in vivo imaging system. Images were acquired in 10° steps over 380°. A rectangular region of interest (ROI) was placed across the full image width at the model disease site. Within the ROI, and for every image, mean fluorescence intensity was computed after background subtraction. In the mouse models studied, the labeled nanovesicles were taken up in both the orthotopic and transgenic brain tumors, and in the arthritic sites (toes and ankles). Curve analysis of the multi angle image ROIs determined the angle with the highest signal. Thus, the optimal angle for imaging each disease site was characterized. The MAROI method applied to imaging of fluorescent compounds is a noninvasive, economical, and precise tool for in vivo quantitative analysis of the disease states in the described mouse models.

The continuous evolution of antimicrobial resistance poses a major threat to public health worldwide. Molecular biology techniques have been integrated to epidemiological surveillance systems to improve the control strategies of this phenomenon.To describe the phenotypic and molecular profiles of the most important Gram negative bacilli from intensive care units in 23 Colombian hospitals during the study period 2009-2012.A descriptive study was conducted in 23 hospitals belonging to the Colombian Nosocomial Resistance Study Group. A total of 38.048 bacterial isolates were analyzed using WHONET over a four-year period. The antimicrobial resistant profiles were described for Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, and Acinetobacter baumannii . Polymerase chain reaction was performed in 1.248 strains to detect the most clinically relevant carbapenemases.Escherichia coli was the most frequently isolated organism (mean=14.8%). Frequency of K. pneumoniae increased significantly from 11% in 2009 to 15% in 2012 (p0.001). All screened isolates had rising trends of multidrug-resistant profiles. KPC ( Klebsiella pneumoniae carbapenemase) was detected in 68.4% of K. pneumoniae isolates while VIM (Verona integron-encoded metallo-betalactamase) was present in 46.5% of them.In this study, an increase in the trend of multidrug-resistant organisms and a wide distribution of carbapenemases was observed. The integration of molecular biology to surveillance systems allowed the compilation of this data, which will aid in the construction of guidelines on antimicrobial stewardship for prevention in Colombia.

Background Recruitment of macrophage precursors to the adventitia plays a key role in the pathogenesis of abdominal aortic aneurysms ( AAA s), but molecular mechanisms remain undefined. The innate immune signaling molecule CD 14 was reported to be upregulated in adventitial macrophages in a murine model of AAA and in monocytes cocultured with aortic adventitial fibroblasts (AoAf) in vitro , concurrent with increased interleukin‐6 (IL‐6) expression. We hypothesized that CD 14 plays a crucial role in adventitial macrophage precursor recruitment early during AAA formation. Methods and Results CD 14 −/− mice were resistant to AAA formation induced by 2 different AAA induction models: aortic elastase infusion and systemic angiotensin II (Ang II ) infusion. CD 14 gene deletion led to reduced aortic macrophage infiltration and diminished elastin degradation. Adventitial monocyte binding to Ang II ‐infused aorta in vitro was dependent on CD 14, and incubation of human acute monocytic leukemia cell line‐1 (THP‐1) monocytes with IL ‐6 or conditioned medium from perivascular adipose tissue ( PVAT ) upregulated CD 14 expression. Conditioned medium from AoAf and PVAT induced CD 14‐dependent monocyte chemotaxis, which was potentiated by IL ‐6. CD 14 expression in aorta and plasma CD 14 levels were increased in AAA patients compared with controls. Conclusions These findings link CD 14 innate immune signaling via a novel IL ‐6 amplification loop to adventitial macrophage precursor recruitment in the pathogenesis of AAA .

Epoxyeicosatrienoic acids (EETs), synthesized and released by astrocytes in response to glutamate, are known to play a pivotal role in neurovascular coupling. In vascular smooth muscle cells (VSMC), EETs activate large-conductance, Ca2+-activated K+(BK) channels resulting in hyperpolarization and vasodilation. However, the functional role and mechanism of action for glial-derived EETs are still to be determined. In this study, we evaluated the effect of the synthetic EET analog 11-nonyloxy-undec-8(Z)-enoic acid (NUD-GA) on outward K+currents mediated by calcium-activated K+channels. Addition of NUD-GA significantly increased intracellular Ca2+and outward K+currents in perivascular astrocytes. NUD-GA-induced currents were significantly inhibited by BK channel blockers paxilline and tetraethylammonium (TEA) (23.4 ± 2.4%; P < 0.0005). Similarly, NUD-GA-induced currents were also significantly inhibited in the presence of the small-conductance Ca2+-activated K+channel inhibitor apamin along with a combination of blockers against glutamate receptors (12.8 ± 2.70%; P < 0.05). No changes in outward currents were observed in the presence of the channel blocker for intermediate-conductance K+channels TRAM-34. Blockade of the endogenous production of EETs with N-methylsulfonyl-6-(2-propargyloxyphenyl)hexanamide (MS-PPOH) significantly blunted ( dl)-1-aminocyclopentane-trans-1,3-dicarboxylic acid ( t-ACPD)-induced outward K+currents ( P < 0.05; n = 6). Both NUD-GA and t-ACPD significantly increased BK channel single open probability; the later was blocked following MS-PPOH incubation. Our data supports the idea that EETs are potent K+channel modulators in cortical perivascular astrocytes and further suggest that these metabolites may participate in NVC by modulating the levels of K+released at the gliovascular space.

This article presents a detector system consisting of three components, a CMOS imaging array, a gaseous-detector structure with a Micromegas layout and a UV-photon sensitive CsI reflective photocathode. All three elements have been monolithically integrated using simple post-processing steps. The Micromegas structure and the CMOS imaging chip are not impacted by the CsI deposition. The detector operated reliably in He/isobutane mixtures and attained charge gains with single photons up to a level of 6 \cdot 10^4. The Timepix CMOS array permitted high resolution imaging of single UV-photons. The system has an MTF50 of 0.4 lp/pixel which corresponds to app. 7 lp/mm., Comment: 4 pages with 8 figures. Submitted to Nucl. Instr. and Meth. A (Elsevier) for proceedings of VCI 2010.

A growing number of studies support an important contribution of astrocytes to neurovascular coupling, i.e., the phenomenon by which variations in neuronal activity trigger localized changes in blood flow that serve to match the metabolic demands of neurons. However, since both constriction and dilations have been observed in brain parenchymal arterioles upon astrocyte stimulation, the specific influences of these cells on the vasculature remain unclear. Using acute brain slices, we present evidence showing that the specific degree of constriction of rat cortical arterioles (vascular tone) is a key determinant of the magnitude and polarity of the diameter changes elicited by signals associated with neurovascular coupling. Thus elevation of extracellular K+ concentration, stimulation of metabotropic glutamate receptors (mGluR), or 11,12-epoxyeicosatrienoic acid application all elicited vascular responses that were affected by the particular resting arteriolar tone. Interestingly, the data suggest that the extent and/or polarity of the vascular responses are influenced by a delimited set point centered between 30 and 40% tone. In addition, we report that distinct, tone-dependent effects on arteriolar diameter occur upon stimulation of mGluR during inhibition of enzymes of the arachidonic acid pathway [i.e., phospholipase A2, cytochrome P-450 (CYP) ω-hydroxylase, CYP epoxygenase, and cycloxygenase-1]. Our findings may reconcile previous evidence in which direct astrocytic stimulation elicited either vasoconstrictions or vasodilations and also suggest the novel concept that, in addition to participating in functional hyperemia, astrocyte-derived signals play a role in adjusting vascular tone to a range where dilator responses are optimal.

Normal brain function requires proper supply of oxygen and glucose in a timely and local manner. This is achieved through an orchestrated intercellular communication between neurones, astrocytes and microvessels that results in a rapid and restricted increase in cerebral blood flow, a process known as neurovascular coupling. Astrocytic end-feet make close contacts with neuronal synapses and blood vessels and, given their ability to release vasoactive signals following neuronal activation, have been recognized as key intermediaries in the neurovascular response. Both dilating and constricting signals appear to be released from astrocytes upon increases in intracellular Ca(2+) concentration, and both dilatation and constriction of brain vessels have been observed in previous studies. In this article, we discuss the various astrocyte-derived vasodilating and vasoconstricting signals, their interactions and effects on astrocytes and vascular smooth muscle cells, and suggest the importance of the intrinsic properties of the latter cell type on the overall neurovascular response. We present a working model in which the rise in astrocytic Ca(2+) following neuronal activation leads not only to the rapid activation of calcium-activated K(+) channels in astrocytic end-feet, but also to their modulation by metabolites of the arachidonic acid pathway, which in general have been proposed to act on vascular smooth muscle cells rather than on astrocytes. We propose that this latter mechanism may in turn modulate K(+) signalling from astrocytes to smooth muscle cells, influencing the overall effects of the vasodilating and vasoconstricting signals released during neuronal activation.

INTRODUCTION: Glioblastoma Multiforme (GBM) is the most aggressive form of primary brain tumor with a median survival of

SapC-DOPS nanovesicles are a stable protein-lipid mixture assembled from saposin C (SapC) and dioleylphosphatidylserine (DOPS). It shows selective tumor-targeting activity in multiple animal tumor models and specific induction of apoptosis in various cancer cell lines. The preferential targeting is due to higher levels of exposed phosphatidylserine on the outer cell membrane of cancer cells. Importantly, induction of apoptosis by SapC-DOPS was shown to occur through intracellular ceramide elevation. Ceramide is known to be a tumor suppressor molecule capable of acting synergistically with Bax and regulating mitochondrial-mediated apoptosis. Effectors that mediate SapC-DOPS-induced apoptosis downstream of ceramide are currently unknown. Therefore, we hypothesized that mitochondria could play a major role during SapC-DOPS induced apoptotic cancer cell death. Cell death following SapC-DOPS treatment was assessed by several indices such as: G6PD release assay to estimate necrosis, DAPI staining and cell cycle analysis to estimate apoptosis and DNA fragmentation, JC-1 assay to estimate mitochondrial membrane potential (ΔΨM) and caspase-3 activation by Western blotting. SapC-DOPS treatment induced significant apoptosis as denoted by the appearance of a subG1 peak, loss of ΔΨM, mitochondrial release of Smac, Cyto c and AIF, as well as mitochondrial Bax translocation and oligomerization. ShRNA-mediated Smac knockdown and Bax inhibition with V5 peptide led to retention of ΔΨM and abrogation of apoptosis with decreased caspase-3 activation. Our results indicate that Smac and Bax play major roles in SapC-DOPS-induced mitochondria-mediated apoptotic pathway in human neuroblastoma cells

Despite significant advances in our understanding of the biology of CNS tumors, the translation of such knowledge to novel and effective therapeutic strategies has been slow. Consequently, both primary (e.g., glioblastoma multiforme) and secondary (metastatic) brain tumors remain among the most intractable and fatal of all cancers. We have shown that nanovesicles consisting of saposin C (SapC) and dioleylphosphatidylserine (DOPS) effectively target and kill brain tumor cells both in vitro and in vivo. These actions are a consequence of the affinity of SapC-DOPS for phosphatidylserine (PS), an acidic phospholipid abundantly present in the outer membrane of a variety of tumor cells and tumor-associated vasculature. Here, we characterize SapC-DOPS bioavailability in a human glioblastoma orthotopic mouse model and reveal a time-dependent, tumor-specific extravascular accumulation of fluorescently labeled SapC-DOPS. Glioblastoma targeting by SapC-DOPS is abrogated after in vivo exposure to lactadherin, a protein that binds PS with high affinity. We also demonstrate that SapC-DOPS selectively targets brain metastases-forming cancer cells both in vitro, in co-cultures with human astrocytes, and in mouse models bearing brain metastases derived from human breast or lung cancer. We finally that SapC-DOPS cytotoxic activity against metastatic breast cancer cells in vitro, and prolongs the survival of mice with breast cancer brain metastases. Taken together, these results suggest that surface-exposed PS can be potentially useful as a novel lipid marker for diagnosis, monitoring, and therapy of central nervous system cancer, and highlights the potential of SapC-DOPS in the diagnosis and treatment of primary and metastatic brain tumors.

Abstract 3198 Background: Red blood cell (RBC) extravasation contributes to the growth of atherosclerotic plaque, and is associated with plaque rupture. High Fat Diet (HFD) fuels systemic inflammation by promoting monocyte activation and their recruitment/transendothelial migration. RBC bind chemokines such as monocyte chemotactic protein-1 (MCP-1) via Duffy Antigen Receptor for Chemokines (DARC). While it was previously reported that HFD may impact RBC cholesterol content, very little is known about the impact – if any – of HFD on RBC function(s). Hence, we studied whether HFD affects biochemical and functional properties of RBC in ways potentially relevant to the progression of atherosclerosis. Methods and Results: Wild type (WT) C56BL6/J mice were fed either chow (10% total fat) or HFD (60% total fat) for 312 weeks. MCP-1 levels were measured in plasma before and after the release of DARC-bound MCP-1 by heparin. Released MCP-1 was 1.5 fold higher in HFD RBC (12 weeks on diet, n=3) compared to chow RBC. Levels of intracellular reactive oxygen species (ROS) were measured by DCFH fluorescence using flow cytometry, and were increased in HFD RBC (17 weeks on diet, n=3) by ∼1.2 fold compared to chow RBC (p Conclusions: We here show that HFD elicits an increase in surface levels of MCP-1 in RBC, which is associated with an enhanced macrophage adhesion to the intact vascular endothelium and increased transendothelial migration in vitro. Further, HFD increases ROS levels and PS externalization in RBC, and HFD RBC exhibit an enhanced splenic uptake in vivo. These effects of HFD on circulating RBC are likely to promote chronic inflammation, rendering RBC a heretofore unknown contributor to local as well as systemic changes that lead to atherosclerosis. Disclosures: No relevant conflicts of interest to declare.