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In a previous work, we identified nine founder mutations present in close to 80% of BRCA1 and BRCA2 mutation carriers, and distributed across the country. The presence of founder mutations constitutes a valuable opportunity to develop new strategies for genetic screening. Genetic tests are primarily performed by NGS sequencing, which requires sophisticated and expensive equipment, and it takes 2-3 weeks for the results to be informed to the patient. In addition, genetic tests are not covered by insurance companies in Latin American countries. In this work, we present the standardization and technical validation of a real-time PCR based methodology for allelic discrimination in order to identify the nine Chilean founder mutations in BRCA1 and BRCA2 genes.We designed nine pairs of probes and nine pairs of primers to amplify synchronically nine regions of the BRCA1/BRCA2 genes by real-time PCR, in order to identify the nine founder mutations through allelic discrimination analyses. Technical validation was performed using 90 positive and 90 negative samples for each mutation. The methodology was tested in a second group of 60 patients. Our method correctly classified carriers and non-carriers of one of the nine Chilean founder mutations with a 100% specificity and 100% sensitivity, compared with Sanger sequencing performance.We develop an inexpensive, simple, and fast mutation detection method that could be implemented locally in Hospitals from the Private to Public health system. This methodology may be useful for the screening of BRCA1 and BRCA2 mutations in other populations.

Background The main objective of this research was to analyze the characteristics of electrical activity in the brain during REM (Rapid Eye Movements) sleep, by using an experimental model a pathology that affects the frontal lobes, such as brain tumors. In addition to determining the impact of variables such as the frontal area (dorsolateral, medial and orbital), laterality and size of the lesion; as well as the demographic and clinical characteristics of the patients evaluated. Methods By using polysomnographic recordings, 10 patients were evaluated. We obtained power spectra through a homemade program. For quantitative EEG (Electroencephalogram) (qEEG) analysis, the Fast Fourier Transform (FFT) algorithm was used to obtain the spectral power of each participant, channel, and frequency band. Results Sleep architecture and spectral power was found to be modified in patients compared to normative values. Other sociodemographic and clinical characteristics of the patients were also influenced, such as age range and antiepileptic drugs. Conclusions Brain tumors in the frontal lobe can modify the rhythmogenesis of REM sleep, possibly due to changes of brain plasticity as an effect of the pathology. In addition to this, through this study we were able to show the association between neuroanatomical and functional changes, on the characteristics of brain electrical activity in patients with frontal brain tumor. Finally, this qEEG analysis technique allows, on the one hand, to deepen the knowledge and relationship between psychophysiological processes and, on the other hand, to be able to guide therapeutic decisions.

Introduction and objectives: arrhythmogenic cardiomyopathy (ACM) is a hereditary heart disease defined by the progressive replacement of the ventricular myocardium with fibroadipose tissue, which can act as a substrate for arrhythmias, sudden death, or even give rise to heart failure (HF). Sudden death is frequently the first manifestation of the disease, particularly among young patients. The aim of this study is to describe a new pathogenic variant in the PKP2 gene. Methods: A descriptive observational study that included 8 initially non-interrelated families with a diagnosis of ACM undergoing follow-up at our HF and Familial Cardiomyopathies Unit, who were carriers of the NM_004572.3:c.775_776insG; p. (Glu259Glyfs*77) variant in PKP2 gene. The genetic testing employed next-generation sequencing for the index cases, and the Sanger method for the targeted study with family members. We compiled personal and family histories, demographic and clinical characteristics, data from the additional tests at the time of diagnosis, and arrhythmic events at diagnosis and during follow-up. Results: We included 47 subjects, of whom 8 were index cases (17%). Among the evaluated family members, 16 (34%) were carriers of the genetic variant, 3 of whom also had a diagnosis of ACM. The majority were women (26 patients; 55.3%), with a mean age on diagnosis of 48.9 ± 18.6 years, and a median follow-up of 39 [24-59] months. Worthy of note are the high incidence of arrhythmic events as the form of presentation and in follow-up (21.5% and 20.9% respectively), and the onset of HF in 25% of the sample. The most frequent ventricular involvements were right (4 patients 16.7%) and biventricular (4 patients 16.7%); we found no statistical differences in any of the variables analysed. Conclusions: This variant is a pathogenic variant of gene PKP2 that has not previously been described and is not present in the control groups associated with ACM. It has incomplete penetrance, a highly variable phenotypic expressivity, and was identified in 8 families of our geographical area in Malaga (Andalusia, Spain), where we can establish a founder effect and describe the clinical and risk characteristics.

The Paramyxoviridae family includes enveloped single-stranded negative-sense RNA viruses such as measles, mumps, human parainfluenza, canine distemper, Hendra, and Nipah viruses, which cause a tremendous global health burden. The ability of paramyxoviral glycoproteins to merge viral and host membranes allows entry of the viral genome into host cells, as well as cell–cell fusion, an important contributor to disease progression. Recent molecular and structural advances in our understanding of the paramyxovirus membrane fusion machinery gave rise to various therapeutic approaches aiming at inhibiting viral infection, spread, and cytopathic effects. These therapeutic approaches include peptide mimics, antibodies, and small molecule inhibitors with various levels of success at inhibiting viral entry, increasing the potential of effective antiviral therapeutic development.

Coxiella burnetii, the causative agent of Q fever, is a small, coccoid, Gram-negative strict intracellular pathogen. One of the most common ways of acquiring Q fever is through inhalation of aerosols containing the bacteria. Because C. burnetii is highly infectious, spreads easily through the air, and is very resistant to environmental conditions, it is considered a biological threat. This paper presents the development and validation of a specific real-time polymerase chain reaction (real-time PCR or qPCR) assay for the detection of C. burnetii, based on the amplification of a fragment of the isocitrate dehydrogenase (icd) encoding gene. This real-time PCR is highly specific, reproducible, and sensitive, allowing the detection of as few as 5 genome equivalents (GEs) of C. burnetii per reaction. The method enables a rapid preliminary differentiation among strains, based on a point mutation at nucleotide 745 of the icd gene. The assay was successfully evaluated in environmental soil samples; a limit of detection of 3 × 104 colony forming units per 0.5 g of soil (∼3 GEs per reaction) was achieved. The newly developed real-time PCR offers a valuable tool for differential detection of C. burnetii strains in environmental soil samples.

Metastasis, the leading cause of cancer-associated deaths, is promoted by transcription factors SNAIL, SLUG, ZEB1 and TWIST through the activation of epithelial-mesenchymal transition (EMT). MicroRNAs can suppress EMT, emerging as candidate molecular biomarkers and novel therapeutic targets. Herein, we evaluated microRNAs downregulated in breast cancer tissues expressing EMT transcription factors, to find new potential regulators of EMT. MiR-30a, miR-1271, miR-196a, miR-202, miR-210, miR-22, miR-331 and miR-34b were validated. Seven microRNAs downregulated luciferase activity through EMT transcription factors 3’UTR, and all microRNAs decreased cell migration, invasion and/or proliferation. In MDA-MB-231 cells, miR-196a and miR-22 decreased endogenous ZEB1 levels, and miR-30a endogenous CCR7 levels. These results suggest that microRNAs studied are novel regulators of EMT through the control of SNAIL, SLUG, ZEB1 and TWIST. They also regulate the metastatic behavior of cancer cells, and may control the development of lymph node metastasis through the regulation of CCR7.Graphical abstract

Severe COVID-19 is associated with epithelial and endothelial barrier dysfunction within the lung as well as in distal organs. While it is appreciated that an exaggerated inflammatory response is associated with barrier dysfunction, the triggers of vascular leak are unclear. Here, we report that cell-intrinsic interactions between the Spike (S) glycoprotein of SARS-CoV-2 and epithelial/endothelial cells are sufficient to induce barrier dysfunction in vitro and vascular leak in vivo, independently of viral replication and the ACE2 receptor. We identify an S-triggered transcriptional response associated with extracellular matrix reorganization and TGF-β signaling. Using genetic knockouts and specific inhibitors, we demonstrate that glycosaminoglycans, integrins, and the TGF-β signaling axis are required for S-mediated barrier dysfunction. Notably, we show that SARS-CoV-2 infection caused leak in vivo, which was reduced by inhibiting integrins. Our findings offer mechanistic insight into SARS-CoV-2-triggered vascular leak, providing a starting point for development of therapies targeting COVID-19.

Purpose To determine the rate of non-invasive follicular thyroid neoplasm with papillary-like nuclear features (NIFTP) in a multi-institutional series from the Iberian Peninsula and describing this NIFTP cohort. Methods Retrospective study of papillary thyroid carcinoma (PTC) or well-differentiated tumours of uncertain malignant potential (WDT-UMP) diagnosed between 2005 and 2015 and measuring >= 5 mm in adult patients from 17 hospitals. Pathological reports were reviewed to determine the cases that fulfil the original criteria of NIFTP and histology was reassessed. Rates were correlated with the number of PTC and its follicular variant (FVPTC) of each institution. Demographic data, histology, management, and follow-up of the reclassified NIFTP cohort were recorded. Results A total of 182 cases with NIFTP criteria were identified: 174/3372 PTC (rate: 5.2%; range: 0-12.1%) and 8/19 WDT-UMP (42.1%). NIFTP rate showed linear correlation with total PTC (p: 0.03) and FVPTC (p: 0.007) identified at each centre. Ultrasound findings were non-suspicious in 60.1%. Fine-needle cytology or core biopsy diagnoses were undetermined in 49.7%. Most patients were treated with total thyroidectomy. No case had nodal disease. Among patients with total thyroidectomy, 89.7% had an excellent response evaluated 1 year after surgery. There were no structural persistence or relapses. Five patients showed residual thyroglobulin after 90 months of mean follow-up. Conclusions NIFTP rate is low but highly variable in neighbouring institutions of the Iberian Peninsula. This study suggests pathologist's interpretation of nuclear alterations as the main cause of these differences. Patients disclosed an excellent outcome, even without using the strictest criteria.

Mindanao adasının kuzeyinde ve Bohol denizinin güneyinde yer alan Camiguin adası Filipinlerin en önemli ve en ilginç 25 turizm merkezinden biridir. Çok sayıda volkanik dağ, lagün gölü, kaplıca, mağara ve ilginç şelale barındıran ada dünyanın en iyi 10 dalgıçlık merkezlerinden biri olarak seçilmiştir. Camiguin adasının en büyük turizm avantajlarından birisi küçük yüzölçümüne sahip olmasına rağmen çok çeşitli turizm olanakları, çok sayıda kaliteli konaklama tesisi ve zengin rekreasyon faaliyetlerini aynı anda sunabilmesidir. Bu çalışmada Filipinler Cumhuriyeti’nde ve Mindanao Adalar Grubu’nda yer alan Camiguin adasının başlıca coğrafi özellikleri ve turizm potansiyeli irdelenmiştir. Bu araştırmada adanın coğrafi konumu, doğal ve beşeri özellikleri tartışıldığı gibi turizm çekicilikleri ve başlıca turizm kaynakları da kısaca ele alınmıştır. Adanın başlıca plajları, kentleri, şelaleleri, mağaraları, volkanları, inanç merkezleri ve tarihsel-kültürel çekicilikleri irdelenerek adanın turizm potansiyeli resmedilmeye çalışılmıştır. Çalışmanın sonuç kısmında, Camiguin adasının en büyük turizm avantajları nelerdir sorusuna yanıt aranmıştır, The Camiguin Island located to the north of the Mindanao Island and to the south of the Bohol Sea, is one of the 25 most significant and interesting tourist destinations in the Philippines. The island, which possesses many volcanic mountains, lagoon lakes, hot springs, caves and interesting waterfalls, has been nominated as one of the top 10 diving centers in the world. One of the major tourism advantages of the Camiguin Island is that despite its small surface area, it offers a wide range of tourism opportunities, ample quality accommodation facilities and rich recreational activities at the same time. In this study, the main geographical features and tourism potential of the Camiguin Island, which is located in the Republic of Philippines and Mindanao Islands Group, have been examined. In the present study, the geographical location of the island, its natural and human characteristics have been discussed; besides, the touristic attractions and major tourism resources have been briefly discussed. We tried to identify the tourism potential of the island by examining the main beaches, cities, waterfalls, caves, volcanoes, belief centers and historical-cultural attractions. In the conclusion part of the study, the answer to the question of what are the biggest tourism advantages of the Camiguin Island has been sought.

Introducción: los trastornos músculo esqueléticos son definidos como problemas de salud asociados con el aparato locomotor, inducidos por el trabajo que se desempeña y las condiciones en las que se ejerce, su principal sintomatología es el dolor, adormecimiento, rigidez, hormigueo, inflamación o pérdida de la fuerza. El profesional de enfermería (PE) realiza actividades en el ámbito laboral que influyen directamente en su aparición. Objetivo: conocer cuáles son los síntomas músculo esqueléticos más frecuentes en el PE en un hospital de segundo nivel. Material y métodos: estudio observacional, descriptivo, prospectivo, transversal, la muestra fue de n= 220 PE, de ambos sexos, de cualquier servicio, de base o contrato, que desempeñan funciones asistenciales. Se utilizó el cuestionario nórdico para recabar la información. Resultados: el 80.9% fueron mujeres, 50% con edad comprendida entre 20 y 39 años, 88.2% manifestó sintomatología durante los últimos 12 meses, 76% manifestó sintomatología los últimos 7 días, las regiones anatómicas más afectadas fueron cuello (41.3%), espalda baja (40.9%), rodillas (37.8%), la mayor frecuencia de sintomatología músculo esquelética se presenta en el turno matutino con 26%. Conclusión: un importante porcentaje de PE manifestó sintomatología músculo esquelética, misma que llegan a interferir con la realización de las actividades normales.

The Paramyxoviridae family comprises important pathogens that include measles (MeV), mumps, parainfluenza, and the emerging deadly zoonotic Nipah virus (NiV) and Hendra virus (HeV). Paramyxoviral entry into cells requires viral-cell membrane fusion, and formation of paramyxoviral pathognomonic syncytia requires cell-cell membrane fusion. Both events are coordinated by intricate interactions between the tetrameric attachment (G/H/HN) and trimeric fusion (F) glycoproteins. We report that receptor binding induces conformational changes in NiV G that expose its stalk domain, which triggers F through a cascade from prefusion to prehairpin intermediate (PHI) to postfusion conformations, executing membrane fusion. To decipher how the NiV G stalk may trigger F, we introduced cysteines along the G stalk to increase tetrameric strength and restrict stalk mobility. While most point mutants displayed near-wild-type levels of cell surface expression and receptor binding, most yielded increased NiV G oligomeric strength, and showed remarkably strong defects in syncytium formation. Furthermore, most of these mutants displayed stronger F/G interactions and significant defects in their ability to trigger F, indicating that NiV G stalk mobility is key to proper F triggering via moderate G/F interactions. Also remarkably, a mutant capable of triggering F and of fusion pore formation yielded little syncytium formation, implicating G or G/F interactions in a late step occurring post fusion pore formation, such as the extensive fusion pore expansion required for syncytium formation. This study uncovers novel mechanisms by which the G stalk and its oligomerization/mobility affect G/F interactions, the triggering of F, and a late fusion pore expansion step-exciting novel findings for paramyxoviral attachment glycoproteins.

SummarySevere COVID-19 is associated with epithelial and endothelial barrier dysfunction within the lung as well as in distal organs. While it is appreciated that an exaggerated inflammatory response is associated with barrier dysfunction, the triggers of this pathology are unclear. Here, we report that cell-intrinsic interactions between the Spike (S) glycoprotein of SARS-CoV-2 and epithelial/endothelial cells are sufficient to trigger barrier dysfunction in vitro and vascular leak in vivo, independently of viral replication and the ACE2 receptor. We identify an S-triggered transcriptional response associated with extracellular matrix reorganization and TGF-β signaling. Using genetic knockouts and specific inhibitors, we demonstrate that glycosaminoglycans, integrins, and the TGF-β signaling axis are required for S-mediated barrier dysfunction. Our findings suggest that S interactions with barrier cells are a contributing factor to COVID-19 disease severity and offer mechanistic insight into SARS-CoV-2 triggered vascular leak, providing a starting point for development of therapies targeting COVID-19 pathogenesis.

In this study, we describe the pathology of Leishmania infantum infection in naturally infected wild Leporidae and compare diagnosis of infection using histopathology, direct fluorescent antibody (DFA) assay, immunofluorescence antibody test (IFAT) and quantitative real-time PCR (qPCR). Tissues were analysed from 52 European rabbits (Oryctolagus cuniculus) and 7 Iberian hares (Lepus granatensis) from the Community of Madrid (Spain). Our results show that L. infantum infection is associated with only minimal histopathological lesions and that L. infantum amastigotes can be detected by DFA assay in all tissues types tested, including skin. These results were confirmed by qPCR on fresh frozen tissues in 13% of rabbits and 100% of hares. However, L. infantum DNA could not be detected by qPCR on paraffin-embedded tissue obtained by laser capture microdissection. Using the DFA assay to diagnose L. infantum, infection may provide further insights into this disease in wild animals and may allow the precise tissue localization of L. infantum, thereby guiding follow-up tests with more accurate qPCR.

This study presents a new bidirectional DC-DC converter with high gains, based on impedance source. The design of the impedance network has been studied in step-up and step-down modes. The most significant innovations of the proposed converter are the linearity of the high gains in step-up and step-down modes with respect to the cycle variations and a full use of the duty cycle, very few passive and semiconductor elements are needed to implement the converter. The switching techniques are very simple, since only the switching of a single transistor in each mode is required. To improve efficiency, the cores of the inductors have been constructed with nanocrystalline material and the resistance of Z-red has been reduced as much as possible. Finally, a laboratory prototype has been implemented.

This paper presents a model to implement the switching technique of a novel isolated DC/DC converter of full-bridge to link fuel cell power output to the distribution system. The implementation was performed on a DSP F2812 of Texas Instruments. It is also proposed a new start-up system (software), to avoid overcurrent on the input side of inverter. This software avoids using an auxiliary circuit for start-up, so reducing costs, improving volume and weight and maintaining the same efficiency. A prototype has been implemented to validate the performance. Finally, comparative experimental results are presented.

Experimental vaccines for the deadly zoonotic Nipah (NiV), Hendra (HeV), and Ebola (EBOV) viruses have focused on targeting individual viruses, although their geographical and bat reservoir host overlaps warrant creation of multivalent vaccines. Here we explored whether replication-incompetent pseudotyped vesicular stomatitis virus (VSV) virions or NiV-based virus-like particles (VLPs) were suitable multivalent vaccine platforms by co-incorporating multiple surface glycoproteins from NiV, HeV, and EBOV onto these virions. We then enhanced the vaccines’ thermotolerance using carbohydrates to enhance applicability in global regions that lack cold-chain infrastructure. Excitingly, in a Syrian hamster model of disease, the VSV multivalent vaccine elicited safe, strong, and protective neutralizing antibody responses against challenge with NiV, HeV, or EBOV. Our study provides proof-of-principle evidence that replication-incompetent multivalent viral particle vaccines are sufficient to provide protection against multiple zoonotic deadly viruses with high pandemic potential.

Cholesterol has been implicated in various viral life cycle steps for different enveloped viruses, including viral entry into host cells, cell-cell fusion, and viral budding from infected cells. Enveloped viruses acquire their membranes from their host cells. Although cholesterol has been associated with the binding and entry of various enveloped viruses into cells, cholesterol’s exact function in the viral-cell membrane fusion process remains largely elusive, particularly for the paramyxoviruses. Furthermore, paramyxoviral fusion occurs at the host cell membrane and is essential for both virus entry (virus-cell fusion) and syncytium formation (cell-cell fusion), central to viral pathogenicity. Nipah virus (NiV) is a deadly member of the Paramyxoviridae family, which also includes Hendra, measles, mumps, human parainfluenza, and various veterinary viruses. The zoonotic NiV causes severe encephalitis, vasculopathy, and respiratory symptoms, leading to a high mortality rate in humans. We used NiV as a model to study the role of membrane cholesterol in paramyxoviral membrane fusion. We used a combination of methyl-beta cyclodextrin (MβCD), lovastatin, and cholesterol to deplete or enrich cell membrane cholesterol outside cytotoxic concentrations. We found that the levels of cellular membrane cholesterol directly correlated with the levels of cell-cell fusion induced. These phenotypes were paralleled using NiV/vesicular stomatitis virus (VSV)-pseudotyped viral infection assays. Remarkably, our mechanistic studies revealed that cholesterol reduces an early F-triggering step but enhances a late fusion pore formation step in the NiV membrane fusion cascade. Thus, our results expand our mechanistic understanding of the paramyxoviral/henipaviral entry and cell-cell fusion processes. IMPORTANCE Cholesterol has been implicated in various steps of the viral life cycle for different enveloped viruses. Nipah virus (NiV) is a highly pathogenic enveloped virus in the Henipavirus genus within the Paramyxoviridae family, capable of causing a high mortality rate in humans and high morbidity in domestic and agriculturally important animals. The role of cholesterol for NiV or the henipaviruses is unknown. Here, we show that the levels of cholesterol influence the levels of NiV-induced cell-cell membrane fusion during syncytium formation and virus-cell membrane fusion during viral entry. Furthermore, the specific role of cholesterol in membrane fusion is not well defined for the paramyxoviruses. We show that the levels of cholesterol affect an early F-triggering step and a late fusion pore formation step during the membrane fusion cascade. Thus, our results expand our mechanistic understanding of the viral entry and cell-cell fusion processes, which may aid the development of antivirals.

Sperm morphology has been used as a prognostic factor in in vitro fertilization, however, in intrauterine insemination (IUI) its predictive role is controversial. The aim of this study was to determine the impact of sperm morphology as isolated parameter of the spermiogram has on the probability of pregnancy in couples that are treated with IUI.Retrospective study of IUI cycles performed in the Reproductive Medicine Unit of Clínica Las Condes between January 2016 and December 2018. Logistic regression of the following variables: sperm morphology grouped in 0-1%, 2-3% and≥4%, total progressively motile sperm count inseminated, age of the woman and the man.A total of 385 cases were included and clinical pregnancy was diagnosed in 85 of them. When separating into groups of sperm morphology4% and≥4%, the pregnancy rate was 22% in both groups. The age of the woman was the only statistically significant factor in the logistic regression. The area under the ROC curve of sperm morphology as a predictor of pregnancy was 0.53.Our study concludes that sperm morphology should not be considered as a single parameter when deciding whether or not a couple can be treated with IUI, eliminating isolated teratozoospermia as a direct indication for in vitro fertilization. It was not possible to determine a cut-off point for sperm morphology that serves as a predictor of pregnancy.

Nipah virus (NiV) is a zoonotic bat henipavirus in the family Paramyxoviridae. NiV is deadly to humans, infecting host cells by direct fusion of the viral and host cell plasma membranes. This membrane fusion process is coordinated by the receptor-binding attachment (G) and fusion (F) glycoproteins. Upon G-receptor binding, F fuses membranes via a cascade that sequentially involves F-triggering, fusion pore formation, and viral or genome entry into cells. Using NiV as an important paramyxoviral model, we identified two novel regions in F that modulate the membrane fusion cascade. For paramyxoviruses and other viral families with class I fusion proteins, the heptad repeat 1 (HR1) and HR2 regions in the fusion protein prefusion conformation bind to form a six-helix bundle in the postfusion conformation. Here, structural comparisons between the F prefusion and postfusion conformations revealed that a short loop region (N1) undergoes dramatic spatial reorganization and a short alpha helix (N4) undergoes secondary structural changes. The roles of the N1 and N4 regions during the membrane fusion cascade, however, remain unknown for henipaviruses and paramyxoviruses. By performing alanine scanning mutagenesis and various functional analyses, we report that specific residues within these regions alter various steps in the membrane fusion cascade. While the N1 region affects early F-triggering, the N4 region affects F-triggering, F thermostability, and extensive fusion pore expansion during syncytium formation, also uncovering a link between F-G interactions and F-triggering. These novel mechanistic roles expand our understanding of henipaviral and paramyxoviral F-triggering, viral entry, and cell-cell fusion (syncytia), a pathognomonic feature of paramyxoviral infections. IMPORTANCE Henipaviruses infect bats, agriculturally important animals, and humans, with high mortality rates approaching ∼75% in humans. Known human outbreaks have been concentrated in Southeast Asia and Australia. Furthermore, about 20 new henipaviral species have been recently discovered in bats, with geographical spans in Asia, Africa, and South America. The development of antiviral therapeutics requires a thorough understanding of the mechanism of viral entry into host cells. In this study, we discovered novel roles of two regions within the fusion protein of the deadly henipavirus NiV. Such roles were in allowing viral entry into host cells and cell-cell fusion, a pathological hallmark of this and other paramyxoviruses. These novel roles were in the previously undescribed N1 and N4 regions within the fusion protein, modulating early and late steps of these important processes of viral infection and henipaviral disease. Notably, this knowledge may apply to other henipaviruses and more broadly to other paramyxoviruses.

As part of the Biology and Mars Experiment (BIOMEX; ILSRA 2009-0834), samples of the lichen Circinaria gyrosa were placed on the exposure platform EXPOSE-R2, on the International Space Station (ISS) and exposed to space and to a Mars-simulated environment for 18 months (2014–2016) to study: (1) resistance to space and Mars-like conditions and (2) biomarkers for use in future space missions (Exo-Mars). When the experiment returned (June 2016), initial analysis showed rapid recovery of photosystem II activity in the samples exposed exclusively to space vacuum and a Mars-like atmosphere. Significantly reduced recovery levels were observed in Sun-exposed samples, and electron and fluorescence microscopy (transmission electron microscope and field emission scanning electron microscope) data indicated that this was attributable to the combined effects of space radiation and space vacuum, as unirradiated samples exhibited less marked morphological changes compared with Sun-exposed samples. Polymerase chain reaction analyses confirmed that there was DNA damage in lichen exposed to harsh space and Mars-like environmental conditions, with ultraviolet radiation combined with space vacuum causing the most damage. These findings contribute to the characterization of space- and Mars-resistant organisms that are relevant to Mars habitability.

Medically important paramyxoviruses, such as measles, mumps, parainfluenza, Nipah, and Hendra viruses, infect host cells by directing fusion of the viral and cellular plasma membranes. Upon infection, paramyxoviruses cause a second type of membrane fusion, cell-cell fusion (syncytium formation), which is linked to pathogenicity. Host cell receptor binding causes conformational changes in the attachment glycoprotein (HN, H, or G) that trigger a conformational cascade in the fusion (F) glycoprotein that mediates membrane fusion. F, a class I fusion protein, contains the archetypal heptad repeat regions 1 (HR1) and 2 (HR2). It is well established that binding of HR1 and HR2 is key to fusing viral and cellular membranes. In this study, we uncovered a novel fusion-modulatory role of a third structurally conserved helical region (HR3) in F. Based on its location within the F structure, and structural differences between its prefusion and postfusion conformations, we hypothesized that the HR3 modulates triggering of the F conformational cascade (still requiring G). We used the deadly Nipah virus (NiV) as an important paramyxoviral model to perform alanine scan mutagenesis and a series of multidisciplinary structural/functional analyses that dissect the various states of the membrane fusion cascade. Remarkably, we found that specific residues within the HR3 modulate not only early F-triggering but also late extensive fusion pore expansion steps in the membrane fusion cascade. Our results characterize these novel fusion-modulatory roles of the F HR3, improving our understanding of the membrane fusion process for NiV and likely for the related Henipavirus genus and possibly Paramyxoviridae family members. IMPORTANCE The Paramyxoviridae family includes important human and animal pathogens, such as measles, mumps, and parainfluenza viruses and the deadly henipaviruses Nipah (NiV) and Hendra (HeV) viruses. Paramyxoviruses infect the respiratory tract and the central nervous system (CNS) and can be highly infectious. Most paramyxoviruses have a limited host range. However, the biosafety level 4 NiV and HeV are highly pathogenic and have a wide mammalian host range. Nipah viral infections result in acute respiratory syndrome and severe encephalitis in humans, leading to 40 to 100% mortality rates. The lack of licensed vaccines or therapeutic approaches against NiV and other important paramyxoviruses underscores the need to understand viral entry mechanisms. In this study, we uncovered a novel role of a third helical region (HR3) of the NiV fusion glycoprotein in the membrane fusion process that leads to viral entry. This discovery sets HR3 as a new candidate target for antiviral strategies for NiV and likely for related viruses.

Glycosylation is a biologically important protein modification process by which a carbohydrate chain is enzymatically added to a protein at a specific amino acid residue. This process plays roles in many cellular functions, including intracellular trafficking, cell–cell signaling, protein folding and receptor binding. While glycosylation is a common host cell process, it is utilized by many pathogens as well. Protein glycosylation is widely employed by viruses for both host invasion and evasion of host immune responses. Thus better understanding of viral glycosylation functions has potential applications for improved antiviral therapeutic and vaccine development. Here, we summarize our current knowledge on the broad biological functions of glycans for the Mononegavirales, an order of enveloped negative-sense single-stranded RNA viruses of high medical importance that includes Ebola, rabies, measles and Nipah viruses. We discuss glycobiological findings by genera in alphabetical order within each of eight Mononegavirales families, namely, the bornaviruses, filoviruses, mymonaviruses, nyamiviruses, paramyxoviruses, pneumoviruses, rhabdoviruses and sunviruses.

The aim of this study was to compare a quantitative real-time PCR (qPCR) validated for the detection of Leishmania infantum in dogs with a nested PCR but in wild Leporidae. Additionally, L. infantum results from indirect immunofluorescent antibody test (IFAT) and in vitro culture were also compared with qPCR. Different samples (spleen, skin and hair) recovered from 224 European rabbits and 70 Iberian hares from two green areas of Madrid Council were analysed for the detection of L. infantum. The presence of Leishmania kDNA was detected by qPCR in 58 out of 221 (26.24%), 162 out of 203 (79.8%) and 22 out of 33 (66.67%) analysed rabbits on spleen, skin and hair samples, respectively; and in 7 out of 69 (10.14%), 39 out of 70 (55.71%) and 17 out of 32 (53.13%) test hares on spleen, skin and hair samples, respectively. The qPCR in all test samples resulted to be more sensitive than nested PCR, with a limit of detection of 1.43 fg/reaction (0.039 parasites) for L. infantum genomic DNA. Additionally, the percentage of positive animals detected by qPCR in at least two out of three samples (n=221 rabbits and 70 hares) tested was higher than those detected by IFAT (n=190 rabbits and 61 hares) and isolation (n=75 rabbits and 20 hares). The highest level of agreement was obtained by nested PCR on spleen/skin (89%/83%) samples and qPCR on spleen samples (81%), followed by IFAT (48%) and qPCR on skin (32%) samples. Our results demonstrate this qPCR is a suitable method for detecting L. infantum DNA in different samples suggesting hair could be considered an adequate sample for direct, reliable and non-invasive diagnosis of L. infantum in wild animals.

Background: Double blockade with pertuzumab and trastuzumab combined with chemotherapy is the standard neoadjuvant treatment for HER2-positive early breast cancer. Data derived from clinical trials indicates that the response rates differ among intrinsic subtypes of breast cancer. The aim of this study is to determine if these results are valid in real-world patients. Methods: A total of 259 patients treated in eight Spanish hospitals were included and divided into two cohorts: Cohort A (132 patients) received trastuzumab plus standard neoadjuvant chemotherapy (NAC), and Cohort B received pertuzumab and trastuzumab plus NAC (122 patients). Pathological complete response (pCR) was defined as the complete disappearance of invasive tumor cells. Assignment of the intrinsic subtype was realized using the research-based PAM50 signature. Results: There were more HER2-enriched tumors in Cohort A (70 vs. 56%) and more basal-like tumors in Cohort B (12 vs. 2%), with similar luminal cases in both cohorts (luminal A 12 vs. 14%; luminal B 14 vs. 18%). The overall pCR rate was 39% in Cohort A and 61% in Cohort B. Better pCR rates with pertuzumab plus trastuzumab than with trastuzumab alone were also observed in all intrinsic subtypes (luminal PAM50 41 vs. 11.4% and HER2-enriched subtype 73.5 vs. 50%) but not in basal-like tumors (53.3 vs. 50%). In multivariate analysis the only significant variables related to pCR in both luminal PAM50 and HER2-enriched subtypes were treatment with pertuzumab plus trastuzumab (Cohort B) and histological grade 3. Conclusions: With data obtained from patients treated in clinical practice, it has been possible to verify that the addition of pertuzumab to trastuzumab and neoadjuvant chemotherapy substantially increases the rate of pCR, especially in the HER2-enriched subtype but also in luminal subtypes, with no apparent benefit in basal-like tumors.

This paper presents a Z-source converter integration technique, with which the energy management of various renewable energy sources is achieved. With a single controller and with serial arrangement of converters, this innovative method connect in series several power sources, with different power and voltage conditions. In this way, the supply to the load can be maintained without alterations, despite the random variations that occur in renewable energy sources. The main innovation is the design of the converters, according to the power and voltage conditions of the sources, so they can be integrated into a common generation branch with a single controller for all converters. The results have been verified by building a laboratory prototype consisting of three unidirectional and two bidirectional converters, which integrate three renewable sources and two electric accumulators. The generation branch management has been implemented in an S7-1500 PLC, each converter has used a DSP TMS320F2812 for the implementation of its switching technique, the branch control has also been implemented in another DSP of the same type.

Supplementary Material The largest phylogenetic lineage known to date of the anthrax pathogen Bacillus anthracis is the wide-spread, so-called Trans-Eurasian clade systematically categorized as the A.Br.008/009 group sharing two defining canonical single-nucleotide polymorphisms (canSNP). In this study, we genome-sequenced a collection of 35 B. anthracis strains of this clade, derived from human infections, animal outbreaks or soil, mostly from European countries isolated between 1936 and 2008. The new data were subjected to comparative chromosomal analysis, together with 75 B. anthracis genomes available in public databases, and the relative placements of these isolates were determined within the global phylogeny of the A.Br.008/009 canSNP group. From this analysis, we have detected 3754 chromosomal SNPs, allowing the assignation of the new chromosomal sequences to established sub-clades, to define new sub-clades, such as two new Spanish, one Bulgarian or one German group(s), or to introduce orphan lineages. SNP-based results were compared with that of a multilocus variable number of tandem repeat analysis (MLVA). This analysis indicated that MLVA typing might provide additional information in cases when genomics yields identical genotypes or shows only minor differences. Introducing the delayed mismatch amplification assay (DMAA) PCR-analysis, we developed a cost-effective method to interrogate for a set of ten phylogenetically informative SNPs within genomes of A.Br.008/009 canSNP clade strains of B. anthracis. By this approach, additional 32 strains could be assigned to five of ten defined clades. View Full-Text This research was funded by funds from the Medical Biological Defense Research Program of the Bundeswehr Joint Medical Service;With funding from the Spanish government through the "María de Maeztu Unit of Excellence" accreditation (MDM-2017-0737). Peer review

A Chilean 35-year-old male patient with a history of primary infertility made an appointment at the Unit of Reproductive Medicine at Clinica Las Condes, Santiago, Chile. Multiple semen analyses revealed abnormal sperm morphology as the most prevalent finding. Multiflagellated and macrocephalic spermatozoa were observed and indicated a possible macrozoospermic phenotype. The constant presence of abnormal sperm morphology led the scope of the study to include Aurora Kinase C (AURKC) gene sequencing. The patient was diagnosed with a homozygous mutation of this gene. The mutation was detected in exon 6, type c.744C>G+/+ (P.Y248*) variant. As previously described in the Human Gene Mutation Database (HGMD), this pathogenic variant is associated with macrozoospermia. Although this mutation is not the most frequently observed, it is the first of its kind reported in Latin America.

BRCA1 has been found to be absent or miss localized in the cytoplasm in a relevant proportion of breast cancer tumors with no germline mutations. BRCA1 main function is in the nucleus, and its interaction with BARD1 is relevant for its nuclear translocation and retention. Our aim was to analyze the sub-cellular localization of BRCA1 and BARD1 in breast cancer tumors, and determine the level of expression of their splice variants BRCA1-Δ11q and BARD1-α and BARD1-β. BRCA1 and BARD1 expressions were performed by immunohistochemistry and immunofluorescence in 103 breast cancer tumors. Colocalization was determined by confocal microscopy. Transcript variants were determined by qRT-PCR. We found BRCA1 localized in the cytoplasm with BARD1 in 51.4 % of tumors. An exclusive nuclear localization of both proteins was observed in 7/103 tumors (6.8 %). Indeed, these tumors displayed an apparent nucleolar colocalization of BARD1 and BRCA1. In relation to splice variants, there is a tendency to an overexpression of BARD1-α mRNA (30 % of tumors) and a decreased expression of BARD1-β (41 %). BRCA1 full-length was downregulated in 63 % of tumors, and 37 % showed BRCA1-Δ11q variant overexpressed. Our findings contribute to a better understanding of the expression and sub-cellular localization of BRCA1 in breast cancer tumors. Interaction of BRCA1 and BARD1 seems to be not affected in 58.2 % of tumors, which showed colocalization of both proteins. The absence of BRCA1 in 41 % of tumors reveals a BRCAness phenotype, constituting an excellent marker for therapy sensitivity, to platinum drugs or PARP inhibitors.

Nipah virus (NiV), a paramyxovirus in the genus Henipavirus , has a mortality rate in humans of approximately 75%. While several studies have begun our understanding of NiV particle formation, the mechanism of this process remains to be fully elucidated. For many paramyxoviruses, M proteins drive viral assembly and egress; however, some paramyxoviral glycoproteins have been reported as important or essential in budding. For NiV the matrix protein (M), the fusion glycoprotein (F) and, to a much lesser extent, the attachment glycoprotein (G) autonomously induce the formation of virus-like particles (VLPs). However, functional interactions between these proteins during assembly and egress remain to be fully understood. Moreover, if the F-driven formation of VLPs occurs through interactions with host cell machinery, the cytoplasmic tail (CT) of F is a likely interactive domain. Therefore, we analyzed NiV F CT deletion and alanine mutants and report that several but not all regions of the F CT are necessary for efficient VLP formation. Two of these regions contain YXXØ or dityrosine motifs previously shown to interact with cellular machinery involved in F endocytosis and transport. Importantly, our results showed that F-driven, M-driven, and M/F-driven viral particle formation enhanced the recruitment of G into VLPs. By identifying key motifs, specific residues, and functional viral protein interactions important for VLP formation, we improve our understanding of the viral assembly/egress process and point to potential interactions with host cell machinery. IMPORTANCE Henipaviruses can cause deadly infections of medical, veterinary, and agricultural importance. With recent discoveries of new henipa-like viruses, understanding the mechanisms by which these viruses reproduce is paramount. We have focused this study on identifying the functional interactions of three Nipah virus proteins during viral assembly and particularly on the role of one of these proteins, the fusion glycoprotein, in the incorporation of other viral proteins into viral particles. By identifying several regions in the fusion glycoprotein that drive viral assembly, we further our understanding of how these viruses assemble and egress from infected cells. The results presented will likely be useful toward designing treatments targeting this aspect of the viral life cycle and for the production of new viral particle-based vaccines.

Resumen La aparicion de un nodulo tiroideo se ha convertido en un hecho cotidiano en la practica clinica diaria. Habitualmente, la adecuada evaluacion del nodulo tiroideo requiere la realizacion de diversas pruebas diagnosticas y multiples citas medicas, con la consiguiente demora diagnostica. La instauracion de una consulta de alta resolucion de nodulo tiroideo evita en gran medida estos inconvenientes, condensando en una unica cita todas las pruebas necesarias para la correcta evaluacion del nodulo tiroideo. En este trabajo revisamos cual debe ser la estructura diagnostica y funcional de una consulta de alta resolucion de nodulo tiroideo.

The preparation of an ITO (Indium Tin Oxide) modified electrode with a mixed valence Co III /Fe II complex is described. The ITO electrode was initially modified with Au nanoparticles by dip coating in order to obtain and ITO/Au surface over which a film of the [ trans- L 14 Co III NCFe II (CN) 5 ]Na (L 14 Co III –Fe II ; L 14 = 6-methyl-1,4,8,11-tetraazacyclotetradecan-6-amine) complex was deposited electrochemically, thus obtaining an ITO/Au/L 14 Co III –Fe II modified electrode. The modified electrode has been characterized by cyclic voltammetry, Raman spectroscopy, AFM and SEM, thus confirming the presence of the mixed valence complex as a modifier species. The electrode prepared shows good stability in aqueous solution and its activity as electrocatalyst in the reduction of nitrites has been evaluated. Cyclic voltammetry experiments run in presence of nitrites show a current enhancement on the ITO/Au/L 14 Co III –Fe II modified electrodes with respect to the ITO/Au initial electrode. After controlled potential electrolysis experiments, analysis of the products indicate the presence of hydroxylamine as the main nitrite reduction species, although small quantities of hydrazine and ammonia where also obtained.

Creativity, understood as a cerebral function that generates products that are both novel and practical, is one factor that allows us to better adapt to our environment. Sexual hormones, meanwhile, have effects on the central nervous system that can modify it, either temporarily or permanently. It has been shown that these two aspects are interrelated in women, as cerebral activity varies with the phases of the menstrual cycle; for example, in performance on memory-related tasks. Thus, the objective of this research was to determine whether changes in verbal and figural creativity occur during the three phases of the cycle: menstrual, follicular and luteal. The study evaluated 28 healthy women and 10 healthy men, all 18-to-25-year-old undergraduate students. Creativity was measured using the scales from the verbal and figural sections of the Torrance Test of Creative Thinking (TTCT), Forms A and B (counterbalanced), in three sessions programmed to coincide with the three phases of the female subjects’ menstrual cycles. Also, the women filled out the Menstrual Distress Questionnaire (MDQ). For the male participants, the three applications were timed to coincide with the phases of the women’s cycles. Though no significant differences were observed in verbal and figural creative thinking among the phases of the menstrual cycle, significant gender differences were seen, as men achieved higher scores than women on some aspects of figural creativity when the latter were in the follicular and luteal phases. The study concludes that differences in the levels of sexual hormones between men and women influence performance on creativity tasks, and that figural creativity proved to be more sensitive to hormonal change. Finally, the factors of Intellectual Quotient (IQ) (evaluated by the WAIS test), and verbal and spatial abilities (assessed using the DAT test), were not found to have any effect on creativity.

Resumen Fundamentos y objetivos El hallazgo de un nodulo tiroideo es un hecho muy frecuente en la practica clinica habitual. El manejo del nodulo tiroideo necesita un abordaje multidisciplinar que puede ser facilitado con la creacion de unidades de alta resolucion de nodulo tiroideo. Describimos la experiencia clinica y los resultados obtenidos en una consulta de alta resolucion y acto unico de nodulo tiroideo. Material y metodos Se han evaluado todos los pacientes derivados a la consulta de nodulo tiroideo del Hospital Virgen de la Victoria (Malaga) desde el ano 2005 al 2007. Se han seguido las guias de practica clinica de nodulo tiroideo para la recomendacion de puncion aspirativa con aguja fina (PAAF) y cirugia tiroidea. Se realizo analisis estadistico con programa SPSS. Resultados Durante el periodo de estudio, la consulta de nodulo tiroideo atendio a 658 pacientes (edad media 48,6 anos, 89,8% mujeres). Mediante ecografia se confirmo la existencia de nodulo tiroideo en el 85,5% de los pacientes. Con un diametro medio de 1,96 cm, un 74,2% de nodulos fueron solidos, un 55,8% hipoecoicos, y un 4,7% presentaron microcalcificaciones. Se realizo la PAAF a 475 pacientes. El 76,5% de las citologias fueron benignas, el 19,1% sospechosas, el 2,1% malignas y el 2,3% insuficientes. Se derivo a cirugia al 23,3% de los pacientes (principalmente por tamano nodular o sospecha de malignidad). Se confirmo la existencia de patologia maligna (principalmente carcinoma papilar) en el 24,9% de los nodulos intervenidos. La sensibilidad/especificidad de la citologia (considerando la biopsia como el gold standard) fue del 81,8 y el 94,7%, respectivamente. Conclusiones La instauracion de una consulta monografica de nodulo tiroideo permite la optimizacion de recursos y reduce la demora diagnostica, obteniendo unos resultados clinicos ampliamente satisfactorios.

Background and objectives The finding of a thyroid nodule is a very common occurrence in clinical practice. Management of thyroid nodules requires a multidisciplinary approach that may be eased by a high-resolution thyroid nodule clinic. We report our clinical experience and outcomes in a high-resolution thyroid nodule clinic. Patients and methods All patients referred to Virgen de la Victoria Hospital (Malaga) from 2005 to 2007 were evaluated following thyroid nodule guidelines. Statistical analysis was performed using SPSS software. Results In the study period, 658 patients (mean age 48.6 years, 89.8% females) were seen at the thyroid nodule clinic. Thyroid nodules were discovered in 85.5% of patients. Mean nodule diameter was 1.96 cm. Of these nodules, 74.2% were solid, 55.8% hypoechogenic, and 4.7% showed microcalcifications. Fine needle aspiration (FNA) was performed in 475 patients (76.5% of all cytological samples were benign, 19.1% had suspected malignancy, 2.1% were malignant, and 2.3% inadequate). Referral for surgery was required in 23.3% of patients (mainly because of nodule size or suspected malignancy). Malignancy was confirmed in 24.9% of nodules. Sensitivity and specificity of cytology (considering biopsy as gold standard) were 81.8% and 94.7% respectively. Conclusions Implementation of a high-resolution thyroid nodule clinic decreases delay in diagnosis and optimizes available resources, thus providing for satisfactory clinical outcomes.

Coated silver (Ag) colloids synthesized with D-glucose permit the observation of surface-enhanced fluorescence (SEF) and surface-enhanced resonance Raman scattering (SERRS) of the rhodamine B (RhB) molecule. The organic coating formed during the synthesis of the Ag nanostructures was identified by its surface-enhanced Raman scattering (SERS) spectrum as D-gluconic acid. The RhB molecule is used to exemplify the distance dependence of SEF and SERRS on the coated Ag nanostructures. The fluorescence enhancement factor for RhB on D-gluconic acid coated silver nanoparticles was determined experimentally and estimated using a simple model. Further support for the plasmon enhancement is obtained from the fact that the measured fluorescence lifetime of RhB on the silver coated with D-gluconic acid is shorter than that found on a glass surface. A very modest enhancement factor is obtained, as expected for very short distance between RhB and the metal surface. Given the very thin metal-fluorophore separation, estimated from the size of the D-gluconic acid, the energy transfer or fluorescence quenching is still efficient and the SEF enhancement is just overcoming the energy transfer. Therefore, both SEF and SERRS are observed. Notably, the aggregation of coated nanoparticles also increases the enhancement factor for SEF.

Diverse studies have concluded that Vitamin D plays a main role of in breast cancer risk. The active metabolite of vitamin D, is a pleiotropic hormone that regulates proliferation, survival, and differentiation of many cell types, through its nuclear receptor VDR. Clinical and epidemiological studies have described the association of decreased serum concentrations of 25-hydroxy-vitamin D with a poor prognosis in patients with breast cancer. The metabolism of vitamin D is meticulously regulated through a complex process, involving the vitamin D-activating enzyme CYP27B1, responsible for the final hydroxylation step to form the active metabolite and CYP24A1, the key enzyme in the inactivation of this metabolite. It is also known that VDR is central for vitamin D-mediated transcription regulation. For all these processes it is necessary that vitamin D enters the cell. In the kidney, internalization of the circulating vitamin D is accomplished by megalin, a membrane receptor for vitamin D, which has not been defined in mammary tissue. Until today reports published in relation to the mechanism of vitamin D uptake and its metabolism are scarce in breast cancer tumors. Our aim was to quantify the expression and to determine the localization of: megalin, VDR, CYP27B1 and CYP24A1 enzymes in breast cancer tumor tissues and normal breast tissue. For this purpose, tissue microarrays of formalin-fixed and paraffin-embedded tumor samples and normal breast tissues were analyzed through immunohistochemistry. This study showed that megalin is localized apically in epithelial cells of normal breast tissue, as previously described in the kidney. On the other hand, in breast cancer tumors megalin expression was diminished. Interestingly, we found that megalin had a different localization in two breast cancer tumor subtypes. In luminal breast cancer tumors, megalin was localized mainly in the cellular membrane whereas in triple negative tumors we observed a cytoplasmic localization. For VDR, CYP27B1 and CYP24A1 our results showed that all these proteins are expressed in normal breast tissues. VDR and CYP24A1 have a heterogeneous expression in relation to tumor subtypes and these are highly expressed in all luminal breast cancer tumors, whereas in triple negative breast cancer subtype its expression was low. Our results suggest that exists an alteration in classical metabolic pathway of vitamin D in the tumor subtypes analyzed of breast cancer. With these results we can suggest that in luminal tumors, which have the better prognosis compared with other subtypes, tumor cells still to respond to vitamin D, while in triple negative subtype, the tumor cells reduce their ability to: uptake, catabolize the active metabolite and respond to vitamin D favoring tumor progression. Citation Format: Gabriela Valarezo, Victoria Ortega-Hernández, Gonzalo Escobar-Massú, Wanda Fernández, María Paz Marzolo, Pilar Carvallo. Vitamin D uptake and metabolism in breast cancer tumors: Differential expression of megalin, VDR, CYP27B1 and CYP24A [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 1440.

Breast cancer is the leading cause of cancer death among women worldwide being distant metastases the main cause of disease. Patients with triple negative breast cancer tumors develop distant metastases earlier compared to luminal tumors. Epithelial-mesenchymal transition (EMT) induced by TGFβ signaling pathways, is the main mechanism to promote metastasis through the expression of the transcription factors TWIST, SNAIL, SLUG and ZEB1. Expression of EMT transcription factors has been studied in cancer cell lines however studies in breast cancer tumors with different subtype have not been reported. In addition, the molecular mechanism that induce EMT program is not known yet. In this study we evaluated the expression of the four transcription factors as well as the state of TGFβ/SMAD, ERK/MAPK and PI3K/AKT pathways in breast cancer cell lines HCC1937 (triple negative) and T47D (luminal) after TGFβ treatment. The same analysis was done by immunohistochemistry in 100 breast cancer tumors with different subtype. After TGFβ treatment T47D showed activation of ERK/MAPK and PI3K/AKT pathways and expression of only two transcription factors: SNAIL and SLUG. HCC1937 cells showed activation of TGFβ/SMAD and ERK/MAPK pathways and expression of TWIST, SLUG and ZEB1 but not SNAIL. Similar results were observed in breast cancer tumors, being TGFβ/SMAD pathway active in the majority of triple negative tumors, whereas an active state of PI3K/AKT was observed in luminal tumors. Active ERK/MAPK pathway was observed in both triple negative and luminal tumors. In relation to expression of transcription factors triple negative tumors with an active state of the TGFβ/SMAD pathway showed expression of transcription factors TWIST, SNAIL and SLUG, whereas those tumors with an active state of ERK/MAPK pathway showed only ZEB1 expression. Luminal tumors with an active PI3K/AKT pathway frequently showed ZEB1 expression. Our results showed that expression of a specific EMT transcription factor induced by TGFβ is related to the tumor subtype and depends on the activated signaling pathway. These results suggest different mechanisms of EMT induction by TGFβ in relation to breast cancer tumor subtype. Citation Format: Victoria Ortega-Hernández, Wanda Fernandez, Pilar Carvallo. Alternative activation of TGFβ signaling pathways in breast cancer tumors with different subtype is related to a differential expression of transcription factors that induce EMT [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2036.

Un experimento de riego de clavel en maceta bajo invernadero-umbráculo fue diseñado para conocer la influencia de la hora de riego sobre el crecimiento, desarrollo y la eficiencia de la utilización del agua. Escuela Técnica superior de Ingeniería Agronómica Universidad Politécnica de Cartagena

Breast cancer is the leading cause of cancer death among women worldwide, being distant metastases the main cause of death. Epithelial to mesenchymal transition (EMT) has been implicated in promoting cancer invasion and metastasis. This process can be induced by TGFβ through the expression of transcription factors: TWIST, SNAIL, SLUG and ZEB1, which expression may be promoted by different signaling pathways such as SMAD2/SMAD3, PI3K/AKT and ERK/MAPK. The expression of EMT transcription factors has been previously studied in several cancer cell lines, although no studies have been performed in breast cancer tumors. In this study, we analyzed by immunohistochemistry the expression of the four transcription factors, and the signal transduction key proteins p-SMAD2 (ser465/467), p-SMAD2L (ser245/250/,255) and p-AKT (ser473), in 107 breast cancer tumors. We found expression of at least one transcription factor in all triple negative tumors (n=24), while only 70% luminal tumors showed expression of at least one of these proteins. Expression of ZEB1, SNAIL and SLUG was observed in tumors with metastases to lymph node, whereas the majority of tumors expressing TWIST were negative for lymph node status. All triple negative tumors with an active state of the signal transduction pathway TGFβ/SMAD showed expression of at least one transcription factor, whereas 65% of luminal tumors with an active state of the same pathway showed no expression of these transcription factors. Active state of PI3K/AKT pathway was observed only for luminal tumors as it has been described. Sixty-three percent of tumors having this pathway active showed expression of at least one transcription factor. In conclusion we found differential expression of EMT transcription factors in relation to lymph node status and a differential active state of signaling pathways in relation to tumoral subtype of breast cancer tumors. Our results suggest a different mechanism of EMT induction by TGFβ in relation to two tumor’s subtypes and lymph node metastasis. Citation Format: Victoria Ortega-Hernandez, Patricia Gajardo-Meneses, Wanda Fernandez, Pilar Carvallo. Alternated activation of TGFβ pathway and differential expression of epithelial to mesenchymal transition transcription factors in relation to lymph node metastasis in two subtypes of breast cancer tumors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 864. doi:10.1158/1538-7445.AM2017-864

The objective was to determine the relationship between the mother's nutritional status and the newborn's gestational characteristics. A sample of 149 women with gestational diabetes was controlled in the High Risk Obstetric Unit of the Medical Specialties Public Health Center before delivery in the maternity ward of the Hospital Clinico Herminda Martín de Chillán, Chile in 2010. Data were obtained from the perinatal clinical history and the newborn's chart. The variables recorded for the mother were nutritional status, type of delivery, number of pregnancies, and metabolic control. Data for the newborn were weight, length, head circumference, and gestational diagnosis. These data were analyzed by ANOVA, Chi-square test, and Multiple Correspondence. Women with a normal nutritional status were multiparous with natural childbirth; the newborn had an adequate gestational age and normal head circumference. On the other hand, maternal obesity was related to a Cesarean; the newborn was large for gestational age and had a larger head circumference. Overweight women were primiparous and the newborn was small for gestational age with a smaller head circumference (p0.01). It was concluded that obesity in women with gestational diabetes explains variables such as type of delivery, number of gestations, and the newborn's diagnosis.

Appearance of a thyroid nodule has become a daily occurrence in clinical practice. Adequate thyroid nodule assessment requires several diagnostic tests and multiple medical appointments, which results in a substantial delay in diagnosis. Implementation of a high-resolution thyroid nodule clinic largely avoids these drawbacks by condensing in a single appointment all tests required for adequate evaluation of thyroid nodule. This paper reviews the diagnostic and functional structure of a high-resolution thyroid nodule clinic.

Patients treated with tamoxifen often experience vaginal bleeding, generally because of benign endometrial lesions or, less often, because of endometrial carcinoma. A rare cause of vaginal bleeding in these patients is breast carcinoma metastasis in an endometrial polyp. We describe a 67-yr-old patient with metastasis of lobular breast carcinoma in an endometrial polyp and papillary serous endometrial carcinoma, both diagnosed simultaneously in a uterine curettage. The coexistence of the 2 cancers was confirmed by pathologic study of the hysterectomy specimen. To our knowledge, this is the first reported case of the synchronous presence of the 2 neoplastic diseases. Endometrial biopsies performed for endometrial disorders in patients who have breast cancer and have received tamoxifen should be studied carefully to avoid underdiagnosis.

To report one case of bilateral kidney pseudotumor due to sarcoidosis in a 75 year-old man with prostatic carcinoma and hypertrophic pachymeningitis.Renal Doppler, body CT, CT-guided renal biopsy, blood analysis including angiotensin converting enzyme (ACE), blood levels and test-treatment (corticosteroid response) were performed.The radiological studies performed showed bilateral kidney masses and pulmonary calcified hilar adenopathies. Blood analysis showed renal failure and increased ACE levels. Renal biopsy showed non-caseating granulomas. Neurological symptoms and renal failure improved with corticosteroid therapy.Bilateral kidney pseudotumor due to sarcoidosis is a rare pathology. Sarcoidosis must be included in the differential diagnosis work up of patients with inflammatory or autoimmune disease and bilateral kidney pseudotumors. Radiological findings of kidney sarcoidosis are quite unspecific. Histological diagnosis with CT guided biopsy or US guided biopsy of kidney masses may be performed.

To test prospectively whether moderate hyperhomocysteinemia and low folate levels could have an influence in the prognosis of 155 patients who presented with an acute coronary syndrome.After a mean follow-up of 13.4+/-7.4 months, patients with low folate levels had higher percentages of cardiovascular death and major cardiovascular events (33% vs. 5%, p0.001; 44% vs. 22%, p0.05) and patients with high homocysteine levels had a higher percentage of major cardiovascular events (31% vs. 14.5%, p0.03). Kaplan-Meier survival estimates analysis showed that patients with low folate levels had a significantly higher probability of cardiovascular death and lower free-of-events survival (log rank statistic: 21.17, p0.001 and 6.59, p=0.01). Patients with high homocysteine levels had a lower free-of-events survival (log rank statistic: 4.95, p=0.02). Different survival multivariate analysis model showed that the presence of low folate levels was an independent predictor of cardiovascular death (hazard ratio 8.85, 95% confidence interval 2.6-29.3, p0.000) and high homocysteine levels was identified as independent predictor of major cardiovascular events (hazard ratio 2.34, 95% confidence interval 1.07-5.12, p0.03).Low folate levels and moderate hyperhomocysteinemia were identified as independent predictors of cardiovascular events in the follow-up.